iACP-DFSRA: identification of anticancer peptides based on a dual-channel fusion strategy of ResCNN and Attention.

Anticancer peptides (ACPs) have been widely applied in the treatment of cancer owing to good safety, rational side effects, and high selectivity. However, the number of ACPs that have been experimentally validated is limited as identification of ACPs is extremely expensive. Hence, accurate and cost-effective identification methods for ACPs are urgently needed. In this work, we proposed a deep learning-based model, named iACP-DFSRA, for ACPs identification. Specifically, we adopted two kinds of sequence embedding technologies, ProtBert_BFD pre-training language model and handcrafted features to encode protein sequences. Then, the LightGBM was used for feature selection, and the selected features were input into ResCNN and Attention mechanism, respectively, to extract local and global features. Finally, the concatenate features were deeply fused by using the Attention mechanism to allow key features to be paid more attention to by the model and make predictions by fully connected layer. The results of 10-fold cross-validation demonstrated that the iACP-DFSRA model delivered improved results in most metrics with Sp of 94.15%, Sn of 95.32%, Acc of 94.74% and MCC of 89.48% compared to the latest AACFlow model. Indeed, the iACP-DFSRA model is the only model with Acc > 90% and MCC > 80% on this independent test dataset. Furthermore, we have further demonstrated the superiority of our model on additional datasets. In addition, t-SNE and SHAP interpretation analysis demonstrated that it is crucial to use two channels for feature extraction and use the Attention mechanism for deep fusion, which helps the iACP-DFSRA to predict ACPs more effectively.

Relationship between weight-adjusted-waist index and all-cause and cardiovascular mortality in individuals with type 2 diabetes.

AIM: To investigate the relationship between the weight-adjusted-waist index (WWI) and all-cause mortality as well as cardiovascular mortality in individuals with type 2 diabetes. METHODS: We used data from the National Health and Nutrition Examination Survey (NHANES) from 1999 to 2018 and the UK Biobank database. Restricted cubic spline curves and Cox proportional hazards models were employed to assess hazard ratios (HRs) and 95% confidence intervals (CIs) for all-cause mortality. RESULTS: In the UK Biobank database, compared with the lowest WWI quartile, the HR for all-cause and cardiovascular death in the highest quartile was 1.846 (95% CI 1.687-2.019) and 2.118 (95% CI 1.783-2.517), respectively, in the fully adjusted model. In the NHANES database, compared with the lowest WWI quartile, the highest quartile had an HR of 1.727 (95% CI 1.378-2.163) for all-cause death and 1.719 (95% CI 1.139-2.595) for cardiovascular death in the fully adjusted model. CONCLUSIONS: Our study indicates that WWI has a long-term synergistic negative impact on all-cause mortality and cardiovascular mortality in individuals with type 2 diabetes. The WWI is an independent predictor of mortality in individuals with type 2 diabetes.

Surface-air exchanges of H2S and SO2 in an urban wetland in eastern China.

Wetlands are widely recognized as hot spots for the emission or deposition of biogenic sulfur gases, including hydrogen sulfur (H2S) and sulfur dioxide (SO2), which significantly affect air quality and climate change. With the expansion of urban wetlands, it is critical to know the roles that urban wetlands played in atmospheric H2S and SO2 budget. In this study, the surface-air exchange fluxes of H2S and SO2 were measured by the Dynamic Flux Chamber (DFC) method in a typical urban wetland in eastern China from Sep 2022 to July 2023. It was found that the urban wetland did not have the expected high H2S emission, might be caused by the relatively high pH value and low sulfate concentration in the soil. Although H2S showed emission in the daytime of spring and summer, an overall H2S flux of -0.04 kg S ha-1 yr-1 was observed throughout the year. Meanwhile, the urban wetland presented a net sink of SO2, with a deposition flux of 0.14 kg S ha-1 yr-1. The negative peaks of SO2 flux corresponded to the suddenly elevated SO2 concentration in the ambient air especially in spring and winter. Through linear fitting of SO2 flux and concentration, the concept of SO2 "compensation point" was proposed. The compensation point is the concentration level at which the observed SO2 flux equals zero. The "compensation point" changed with the season and was related to temperature and humidity. The "compensation point" in summer and autumn were larger, being 2.37 ppb and 1.40 ppb, respectively, while they were 1.07 ppb and 0.86 ppb in spring and winter respectively. Our results suggest that the urban wetland expansion may have little risk of increasing air H2S but could act as a significant sink of SO2 with high SO2 concentration in the urban region.

Association between all-cause mortality and vascular complications in U.S. adults with newly diagnosed type 2 diabetes (NHANES 1999-2018).

AIMS: The impact of macrovascular and microvascular complications, the common vascular complications of type 2 diabetes, on long-term mortality has been well evaluated, but the impact of different complications of newly diagnosed type 2 diabetes (diagnosed within the past 2 years) on long-term mortality has not been reported. We aimed to investigate the relationship between all-cause mortality and vascular complications in U.S. adults (aged ≥ 20 years) with newly diagnosed type 2 diabetes. METHODS: We used data from the 1999-2018 National Health and Nutritional Examination Surveys (NHANES). Cox proportional hazard models was used to assess hazard ratios (HR) and 95% confidence intervals for all-cause mortality. RESULTS: A total of 928 participants were enrolled in this study. At a mean follow-up of 10.8 years, 181 individuals died. In the fully adjusted model, the hazard ratio (HR) (95% confidence interval [CI]) of all-cause mortality for individuals with any single complication compared with those with newly diagnosed type 2 diabetes without complications was 2.24 (1.37, 3.69), and for individuals with two or more complications was 5.34 (3.01, 9.46).Co-existing Chronic kidney disease (CKD) and diabetic retinopathy (DR) at baseline were associated with the highest risk of death (HR 6.07[2.92-12.62]), followed by CKD and cardiovascular disease (CVD) (HR 4.98[2.79-8.89]) and CVD and DR (HR 4.58 [1.98-10.57]). CONCLUSION: The presence of single and combined diabetes complications exerts a long-term synergistic adverse impact on overall mortality in newly diagnosed U.S. adults with type 2 diabetes, underscoring the importance of comprehensive complication screening to enhance risk stratification and treatment.

Construction of biochar-based organohalide-respiring bacterial agent for remediation of 2,4,6-trichlorophenol contaminated soil.

Construction of an efficient bio-reductive dechlorination system remains challenging due to the narrow ecological niche and low-growth rate of organohalide-respiring bacteria during field remediation. In this study, a biochar-based organohalide-respiring bacterial agent was obtained, and its performance and effects on indigenous microbial composition, diversity, and inter-relationship in soil were investigated. A well-performing material, Triton X-100 modified biochar (BC600-TX100), was found to have the superior average pore size, specific surface area and hydrophicity, compared to other materials. Interestingly, Pseudomonas aeruginosa CP-1, which is capable of 2,4,6-TCP dechlorination, showed a 348 times higher colonization cell number on BC600-TX100 than that of BC600 after 7 d. Meanwhile, the dechlorination rate in soil showed the highest (0.732 d-1) in the BC600-TX100 bacterial agent than in the other agents. The long-term performance of the BC600-TX100 OHRB agent was also verified, with a stable dechlorination activity over six cycles. Soil microbial community analysis found the addition of the BC600-TX100 OHRB agent significantly increased the relative abundance of genus Pseudomonas from 1.53 % to 11.2 %, and Pseudomonas formed a close interaction relationship with indigenous microorganisms, creating a micro-ecological environment conducive to reductive dechlorination. This study provides a feasible bacterial agent for the in-situ bioremediation of soil contaminated organohalides. ENVIRONMENTAL IMPLICATION: Halogenated organic compounds are a type of toxic, refractory, and bio-accumulative persistent compounds widely existed in environment, widely detected in the air, water, and soil. In this study, we provide a feasible bacterial agent for the in-situ bioremediation of soil contaminated halogenated organic compounds. The application of biochar provides new insights for "Turning waste into treasure", which meets with the concept of green chemistry.

Exploring efficient strategies for air quality improvement in China based on its regional characteristics and interannual evolution of PM2.5 pollution.

Fine particulate matter (PM2.5) harms human health and hinders normal human life. Considering the serious complexity and obvious regional characteristics of PM2.5 pollution, it is urgent to fill in the comprehensive overview of regional characteristics and interannual evolution of PM2.5. This review studied the PM2.5 pollution in six typical areas between 2014 and 2022 based on the data published by the Chinese government and nearly 120 relevant literature. We analyzed and compared the characteristics of interannual and quarterly changes of PM2.5 concentration. The Beijing-Tianjin-Hebei region (BTH), Yangtze River Delta (YRD) and Pearl River Delta (PRD) made remarkable progress in improving PM2.5 pollution, while Fenwei Plain (FWP), Sichuan Basin (SCB) and Northeast Plain (NEP) were slightly inferior mainly due to the relatively lower level of economic development. It was found that the annual average PM2.5 concentration change versus year curves in the three areas with better pollution control conditions can be merged into a smooth curve. Importantly, this can be fitted for the accurate evaluation of each area and provide reliable prediction of its future evolution. In addition, we analyzed the factors affecting the PM2.5 in each area and summarize the causes of air pollution in China. They included primary emission, secondary generation, regional transmission, as well as unfavorable air dispersion conditions. We also suggested that the PM2.5 pollution control should target specific industries and periods, and further research need to be carried out on the process of secondary production. The results provided useful assistance such as effect prediction and strategy guidance for PM2.5 pollution control in Chinese backward areas.

A generalizable reactive blending strategy to construct flame-retardant, mechanically-strong and toughened poly(L-lactic acid) bioplastics.

Poly(L-lactic acid) (PLA) is an environmentally-friendly bioplastic with high mechanical strength, but suffers from inherent flammability and poor toughness. Many tougheners have been reported for PLA, but their synthesis usually involves organic solvents, and they tend to dramatically reduce the mechanical strength and cannot settle the flammability matter. Herein, we develop strong, tough, and flame-retardant PLA composites by reactive blending PLA, 6-((double (2-hydroxyethyl) amino) methyl) dibenzo [c, e] [1,2] oxyphosphate acid 6-oxide (DHDP) and diphenylmethane diisocyanate (MDI) and define it PLA/xGH, where x indicates that the molar ratio of -NCO group in MDI to -OH group in PLA and DHDP is 1.0x: 1. This fabrication requires no solvents. PLA/2GH with a -NCO/-OH molar ratio of 1.02: 1 maintains high tensile strength of 63.0 MPa and achieves a 23.4 % increase in impact strength compared to PLA due to the incorporation of rigid polyurethane chain segment. The vertical combustion (UL-94) classification and limiting oxygen index (LOI) of PLA/2GH reaches V-0 and 29.8 %, respectively, because DHDP and MDI function in gas and condensed phases. This study displays a generalizable strategy to create flame-retardant bioplastics with great mechanical performances by the in-situ formation of P/N-containing polyurethane segment within PLA.

Molecular and Biological Characteristics of a Peach Latent Mosaic Viroid PC Isolate in Peach from China: Base Mutations in Hairpin Stems and Implications for Symptomatology.

Peach latent mosaic viroid (PLMVd) infects peach trees in China and induces a conspicuous albino phenotype (peach calico, PC) that is closely associated with variants containing a 12-to-14 nucleotide hairpin insertion capped by a U-rich loop. Initially, PC disease distribution was limited to parts of Italy, and it was first detected in the field in China in 2019. To explore the molecular and biological characteristics of PLMVd PC isolates in peach in China, we conducted a comprehensive analysis of disease phenotype development and investigated the data-associated pathogenicity and in vivo dynamics of the Chinese isolate PC-A2 using slash-inoculation into GF-305 peach seedlings. Inoculated seedlings displayed PC symptoms much earlier following topping treatment, and PLMVd infectivity was further assessed using bioassay and semiquantitative RT-PCR experiments. Evolutionary analysis showed that the PC isolate and its progeny variants clustered into a single phylogroup distinct from reference PC-C40 isolates from Italy and PC-K1 and PC-K2 from South Korea. Some PC-A2 progeny variants from green leaves of PC-expressing seedlings showed unbalanced point mutations in hairpin stems compared with the PC-C40 reference sequence and constituted a new stem insertion type. The results reveal associations between the recessive phenotypes of peach albino symptoms and base variation in hairpin stem insertions relative to the PC-C40/chloroplastic heat shock protein 90 reference sequence.

Sesquiterpenes from two Compositae plants as promising inhibitors to nuclear hormone receptor 3 of Tribolium castaneum.

Essential oils (EOs) and their volatile secondary metabolites have been proved to be effective on storage pests control, while restricted on the application due to unclear mechanism. Molecular dynamics (MD) simulations and binding free energies analysis provided an effective approach to reveal mechanism on conformational calculation. In this work, the insecticidal and repellent capacities of Praxelis clematidea and Ageratum houstonianum oils and their main components identified by gas chromatography-mass spectrometry (GC-MS) were scientifically measured. Interestingly, P. clematidea oil exhibited strong fumigant toxicity against Tribolium castaneum (LC50 = 7.07 mg/L air). Moreover, two EOs exhibited over 80% repellent rate against T. castaneum at the highest concentration of 78.63 nL/cm2. Furthermore, hundreds of enzymes related to the regulation of biological processes of T. castaneum were screened to explore the underlying molecular mechanism and develop promising insecticides. Besides, top hits were subjected to MD simulations and binding free energies analysis to elucidate complex inter-molecular stability and affinity over simulated time. The results demonstrated that isolongifolene, δ-cadinene, ß-caryophyllene and caryophyllene oxide were prioritized as they were establishing conserved and stable interactions with residues of nuclear hormone receptor 3 (TcHR3) of T. castaneum, which suggested that the four sesquiterpenes have potential to be promising insecticides on storage pests control.

On-chip Y-junction with adaptive power splitting toward ultrabroad bandwidth.

Growing research interests have been directed to the emerging optical communication band at 2-µm wavelengths. The silicon photonic components are highly desired to operate over a broad bandwidth covering both C-band and the emerging 2-µm wave band. However, the dispersions of the silicon waveguides eventually limit the optical bandwidth of the silicon photonic devices. Here, we introduce a topology-optimized Y-junction with a shallow-etched trench and its utility to reverse the detrimental dispersion effect. The shallow trench enables the Y-junction to have an adaptive splitting capability over a broad spectral range. The 0.2-dB bandwidth of the power splitter exceeds 800 nm from 1400 nm to 2200 nm. The device has a compact footprint of 3 µm × 1.64 µm. The device is characterized at the C-band and 2-µm band with a measured excess loss below 0.4 dB for a proof-of-concept demonstration.

Outdoor Health Risk of Atmospheric Particulate Matter at Night in Xi'an, Northwestern China.

The deterioration of air quality via anthropogenic activities during the night period has been deemed a serious concern among the scientific community. Thereby, we explored the outdoor particulate matter (PM) concentration and the contributions from various sources during the day and night in winter and spring 2021 in a megacity, northwestern China. The results revealed that the changes in chemical compositions of PM and sources (motor vehicles, industrial emissions, coal combustion) at night lead to substantial PM toxicity, oxidative potential (OP), and OP/PM per unit mass, indicating high oxidative toxicity and exposure risk at nighttime. Furthermore, higher environmentally persistent free radical (EPFR) concentration and its significant correlation with OP were observed, suggesting that EPFRs cause reactive oxygen species (ROS) formation. Moreover, the noncarcinogenic and carcinogenic risks were systematically explained and spatialized to children and adults, highlighting intensified hotspots to epidemiological researchers. This better understanding of day-night-based PM formation pathways and their hazardous impact will assist to guide measures to diminish the toxicity of PM and reduce the disease led by air pollution.

Local radiotherapy for murine breast cancer increases risk of metastasis by promoting the recruitment of M-MDSCs in lung.

BACKGROUND: Radiotherapy is one of the effective methods for treatment of breast cancer; however, controversies still exist with respect to radiotherapy for patients with TNBC. Here, we intend to explore the mechanism by which local radiotherapy promotes the recruitment of M-MDSCs in the lung and increases the risk of lung metastasis in TNBC tumor-bearing mice. METHODS: A single dose of 20 Gy X-ray was used to locally irradiate the primary tumor of 4T1 tumor-bearing mice. Tumor growth, the number of pulmonary metastatic nodules, and the frequency of MDSCs were monitored in the mice. Antibody microarray and ELISA methods were used to analyze the cytokines in exosomes released by irradiated (IR) or non-IR 4T1 cells. The effects of the exosomes on recruitment of MDSCs and colonization of 4T1 cells in the lung of normal BALB/c mice were observed with the methods of FCM and pathological section staining. T lymphocytes or 4T1 cells co-cultured with MDSCs were performed to demonstrate the inhibitory effect on T lymphocytes or accelerative migration effect on 4T1 cells. Finally, a series of in vitro experiments demonstrated how the exosomes promote the recruitment of M-MDSCs in lung of mice. RESULTS: Even though radiotherapy reduced the burden of primary tumors and larger lung metastatic nodules (≥ 0.4 mm2), the number of smaller metastases (< 0.4 mm2) significantly increased. Consistently, radiotherapy markedly potentiated M-MDSCs and decreased PMN-MDSCs recruitment to lung of tumor-bearing mice. Moreover, the frequency of M-MDSCs of lung was positively correlated with the number of lung metastatic nodules. Further, M-MDSCs markedly inhibited T cell function, while there was no difference between M-MDSCs and PMN-MDSCs in promoting 4T1 cell migration. X-ray irradiation promoted the release of G-CSF, GM-CSF and CXCl1-rich exosomes, and facilitated the migration of M-MDSCs and PMN-MDSCs into the lung through CXCL1/CXCR2 signaling. While irradiated mouse lung extracts or ir/4T1-exo treated macrophage culture medium showed obvious selective chemotaxis to M-MDSCs. Mechanistically, ir/4T1-exo induce macrophage to produce GM-CSF, which further promoted CCL2 release in an autocrine manner to recruit M-MDSCs via CCL2/CCR2 axis. CONCLUSIONS: Our work has identified an undesired effect of radiotherapy that may promote immunosuppressive premetastatic niches formation by recruiting M-MDSCs to lung. Further studies on radiotherapy combined CXCR2 or CCR2 signals inhibitors were necessary.

Two-dimensional semiconducting SnP2Se6 with giant second-harmonic-generation for monolithic on-chip electronic-photonic integration.

Two-dimensional (2D) layered semiconductors with nonlinear optical (NLO) properties hold great promise to address the growing demand of multifunction integration in electronic-photonic integrated circuits (EPICs). However, electronic-photonic co-design with 2D NLO semiconductors for on-chip telecommunication is limited by their essential shortcomings in terms of unsatisfactory optoelectronic properties, odd-even layer-dependent NLO activity and low NLO susceptibility in telecom band. Here we report the synthesis of 2D SnP2Se6, a van der Waals NLO semiconductor exhibiting strong odd-even layer-independent second harmonic generation (SHG) activity at 1550 nm and pronounced photosensitivity under visible light. The combination of 2D SnP2Se6 with a SiN photonic platform enables the chip-level multifunction integration for EPICs. The hybrid device not only features efficient on-chip SHG process for optical modulation, but also allows the telecom-band photodetection relying on the upconversion of wavelength from 1560 to 780 nm. Our finding offers alternative opportunities for the collaborative design of EPICs.

Copper-induced injectable hydrogel with nitric oxide for enhanced immunotherapy by amplifying immunogenic cell death and regulating cancer associated fibroblasts.

BACKGROUND: Immunogenic cell death (ICD) induced by different cancer treatments has been widely evaluated to recruit immune cells and trigger the specific antitumor immunity. However, cancer associated fibroblasts (CAFs) can hinder the invasion of immune cells and polarize the recruited monocytes to M2-type macrophages, which greatly restrict the efficacy of immunotherapy (IT). METHODS: In this study, an injectable hydrogel induced by copper (Cu) has been designed to contain antibody of PD-L1 and nitric oxide (NO) donor. The therapeutic efficacy of hydrogel was studied in 4T1 cells and CAFs in vitro and 4T1 tumor-bearing mice in vivo. The immune effects on cytotoxic T lymphocytes, dendritic cells (DCs) and macrophages were analyzed by flow cytometry. Enzyme-linked immunosorbent assay, immunofluorescence and transcriptome analyses were also performed to evaluate the underlying mechanism. RESULTS: Due to the absorbance of Cu with the near-infrared laser irradiation, the injectable hydrogel exhibits persistent photothermal effect to kill cancer cells. In addition, the Cu of hydrogel shows the Fenton-like reaction to produce reactive oxygen species as chemodynamic therapy, thereby enhancing cancer treatment and amplifying ICD. More interestingly, we have found that the released NO can significantly increase depletion of CAFs and reduce the proportion of M2-type macrophages in vitro. Furthermore, due to the amplify of ICD, injectable hydrogel can effectively increase the infiltration of immune cells and reverse the immunosuppressive tumor microenvironment (TME) by regulating CAFs to enhance the therapeutic efficacy of anti-PD-L1 in vivo. CONCLUSIONS: The ion induced self-assembled hydrogel with NO could enhance immunotherapy via amplifying ICD and regulating CAFs. It provides a novel strategy to provoke a robust antitumor immune response for clinical cancer immunotherapy.

Folded Digital Meta-Lenses for on-Chip Spectrometer.

In-plane diffractive optical networks based on meta-surfaces are promising for on-chip application. The design constraints of regular antenna unit place ultimate limits on the functionalities of the meta-systems. This fundamental limitation has been reflected by the large footprints of cascaded meta-surfaces. Here, we propose a digital meta-lens with a large degree of design freedom, enabling significantly improved beam focusing, collimation, and deflection capabilities. A highly dispersive and compact diffractive optical system is constructed for spectrometer via five layers of meta-lenses in a folded configuration. The device only occupies a 100 µm × 100 µm chip area on a silicon photonic platform. Sparse and continuous spectra reconstruction is achieved over a 35 nm bandwidth. Fine spectral lines separated by 0.14 nm are resolved. In addition to such a compact and high-resolution on-chip spectrometer, it is also expected to be promising for imaging, optical computing, and other applications due to the great versatility of the digital lens design.

Electromyogram-Based Lip-Reading via Unobtrusive Dry Electrodes and Machine Learning Methods.

Lip-reading provides an effective speech communication interface for people with voice disorders and for intuitive human-machine interactions. Existing systems are generally challenged by bulkiness, obtrusiveness, and poor robustness against environmental interferences. The lack of a truly natural and unobtrusive system for converting lip movements to speech precludes the continuous use and wide-scale deployment of such devices. Here, the design of a hardware-software architecture to capture, analyze, and interpret lip movements associated with either normal or silent speech is presented. The system can recognize different and similar visemes. It is robust in a noisy or dark environment. Self-adhesive, skin-conformable, and semi-transparent dry electrodes are developed to track high-fidelity speech-relevant electromyogram signals without impeding daily activities. The resulting skin-like sensors can form seamless contact with the curvilinear and dynamic surfaces of the skin, which is crucial for a high signal-to-noise ratio and minimal interference. Machine learning algorithms are employed to decode electromyogram signals and convert them to spoken words. Finally, the applications of the developed lip-reading system in augmented reality and medical service are demonstrated, which illustrate the great potential in immersive interaction and healthcare applications.

Investigations on the thermoelectric and thermodynamic properties of Y2CT2 (T = O, F, OH).

Using the first-principle calculations combined with the Boltzmann transport theory, we studied the thermoelectric properties of Y2CT2 (T = O, F, OH) MXenes. Specifically, the Seebeck coefficient, thermal and electrical conductivities under constant relaxation time approximation were calculated. Results show that for p-type carriers, Y2CO2 has the largest power factor of up to 0.0017 W m-1 K-2 when the carrier concentration is 4.067 × 1013 cm-2 at 900 K, at the same temperature, for n-type carriers, the concentration is 9.376 × 1013 cm-2, the power factor in Y2C(OH)2 is 0.0026 W m-1 K-2. In particular, the figure of merit in Y2CF2 is 1.38 at 900 K because of its low thermal conductivity, indicating that it can be considered a potential medium-temperature thermoelectric material. In addition, the thermodynamics properties within 32 GPa and 900 K, such as bulk modulus, heat capacity and thermal expansion, are also estimated using the quasi-harmonic Debye model. Our results may offer some valuable hints for the potential application of Y2CT2 (T = O, F, OH) in the thermoelectric field.

Incorporation of the YRHQ motif into CD3ζ chain enhances the antitumor activity of HER2-targeted CAR-T cells / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨在靶向HER2的CAR的CD3ζ链胞内区引入YRHQ基序对CAR-T细胞的特异性杀伤活性及免疫记忆形成的影响。方法：通过DNA合成获得包含靶向HER2的编码抗原受体H28ζ或H28ζ(YRHQ)的DNA片段，通过慢病毒载体将不同CAR的DNA片段分别转导健康人外周血T细胞，制备靶向HER2的H28ζ-CAR-T及H28ζ(YRHQ)-CAR-T细胞。扩增过程中对不同CAR-T细胞进行计数，FCM检测CAR的表达率。将CAR-T细胞分别与HER2阳性的SKOV3、MDA-MB-453或HER2阴性的MCF-7细胞共培养，LDH释放法检测其杀伤活性，ELISA法检测IL-2、IFN-γ和颗粒酶B的水平，WB法检测STAT3磷酸化水平及免疫检查点分子TIM-3和PD-1的表达，通过FCM检测CCR7、CD45RO的表达，分析CAR-T细胞的表型。结果：H28ζ-CAR-T和H28ζ(YRHQ)-CAR-T细胞扩增能力较好，体外培养7 d时扩增4~5倍。H28ζ-CAR和H28ζ(YRHQ)-CAR表达率分别为（33.3±2.85）%和（28.30±3.2）%。H28ξ(YRHQ)-CAR-T细胞的杀伤活性较H28ζ-CAR-T细胞更高（P<0.05）。经HER2抗原刺激后，与T细胞或H28z-CAR-T细胞比较，H28ξ(YRHQ)-CAR-T细胞的STAT3磷酸化水平较H28ξ-CAR-T细胞明显升高（P<0.01）；而两者间PD-1和TIM-3的表达无明显差异。未经抗原刺激的CAR-T细胞CCR7和CD45RO表达与正常T细胞比较差异无统计学意义（均P>0.05），与SKOV3细胞共培养后，与T细胞或H28z-CAR-T细胞比较，H28ξ(YRHQ)-CAR-T细胞中TEM细胞比例明显增加、TCM细胞比例明显减少（均P<0.05）。结论：在CD3胞内区引入YRHQ基序可在一定程度上提高CAR-T细胞的杀伤潜力。

Study on the material basis of Dahuang Zhechong pill of anti-hepatoma effect by promoting vascular normalization.

Dahuang Zhechong Pill (DHZCP) is a traditional Chinese medicine prescription used to treat many diseases especially chronic liver disease accompanied by promotion of vascular normalization. In this work, UPLC-Q-TOF-MS/MS analysis was applied to identify the chemical components absorbed in the blood. HIF-1α, VEGF, Ang2 and Tie2 related to vascular normalization were detected to determine the dynamic changes of pharmacodynamic indicators. Then, the spectrum-effect relationship between the UHPLC fingerprint and pharmacodynamic indicators was evaluated dynamically using partial least squares (PLS). As a result, 103 components were identified from rat serum samples, including 56 original compounds and 47 metabolites. According to the PLS, active constituents of DHZCP acting on HIF-1α, VEGF, Ang2 and Tie2 (8, 15, 17 and 20) were found. In subsequent experiments on cells, 7/11 components of HIF-1α/VEGF were found in HepG2 and HUVEC cells, and 11/14/2 components of HIF-1α/VEGF/Tie2. The main pharmacodynamic components of DHZCP in promoting vascular normalization were successfully identified by the spectrum-effect relationship analysis.