Methodological Validation of Sedimentation Velocity Analytical Ultracentrifugation Method for Adeno-Associated Virus and Collaborative Calibration of System Suitability Substance.

Currently, adeno-associated virus (AAV) is one of the primary gene delivery vectors in gene therapy, facilitating long-term in vivo gene expression. Despite being imperative, it is incredibly challenging to precisely assess AAV particle distribution according to the sedimentation coefficient and identify impurities related to capsid structures. This study performed the systematic methodological validation of quantifying the AAV empty and full capsid ratio. This includes specificity, accuracy, precision, linearity, and parameter variables involving the sedimentation velocity analytical ultracentrifugation (SV-AUC) method. Specifically, SV-AUC differentiated among the empty, partial, full, and high sedimentation coefficient substance (HSCS) AAV particles while evaluating their sedimentation heterogeneity. The intermediate precision analysis of HE (high percentage of empty capsid) and HF (high percentage of full capsid) samples revealed that the specific species percentage, such as empty or full, was more significant than 50%. Moreover, the relative standard deviation (RSD) could be within 5%. Even for empty or partially less than 15%, the RSD could be within 10%. The accuracy recovery rates of empty capsid were between 103.9% and 108.7% across three different mixtures. When the measured percentage of specific species was more significant than 14%, the recovery rate was between 77.9% and 106.6%. Linearity analysis revealed an excellent linear correlation between the empty, partial, and full in the HE samples. The AAV samples with as low as 7.4 × 1011 cp/mL AAV could be accurately quantified with SV-AUC. The parameter variable analyses revealed that variations in cell alignment significantly affected the overall results. Still, the detection wavelength of 235 nm slightly influenced the empty, partial, and full percentages. Minor detection wavelength changes showed no impact on the sedimentation coefficient of these species. However, the temperature affected the measured sedimentation coefficient. These results validated the SV-AUC method to quantify AAV. This study provides solutions to AAV empty and full capsid ratio quantification challenges and the subsequent basis for calibrating the AAV empty capsid system suitability substance. Because of the AAV structure and potential variability complexity in detection, we jointly calibrated empty capsid system suitability substance with three laboratories to accurately detect the quantitative AAV empty and full capsid ratio. The empty capsid system suitability substance could be used as an external reference to measure the performance of the instrument. The results could be compared with multiple QC (quality control) laboratories based on the AAV vector and calibration accuracy. This is crucial for AUC to be used for QC release and promote gene therapy research worldwide.

Corrigendum: Study of quality of life and its correlated factors in patients after lumbar fusion for lumbar degenerative disc disease.

[This corrects the article DOI: 10.3389/fsurg.2022.939591.].

Metastatic Pulmonary Desmoplastic Small Round Cell Tumor on FDG PET/CT.

ABSTRACT: A 33-year-old woman presents with cough and hemoptysis for 1 month. Chest CT showed a soft tissue mass in the left upper lobe of the lung. FDG PET/CT showed multiple foci of intense activity not only in the lung but also in the lymph nodes and the bones, which was diagnosed as lung malignancy with metastases. Histopathology revealed desmoplastic small round cell tumor. Our case indicated that, although the incidence is low, desmoplastic small round cell tumor should be considered among differential diagnoses of lung malignancies.

Quality and consistency of clinical practice guideline recommendations for PET/CT and PET: a systematic appraisal.

OBJECTIVES: To systematically appraise the methodologies used for guidelines for positron emission tomography (PET) imaging and to compare the consistency of these recommendations. METHODS: We searched PubMed, EMBASE, four guideline databases, and Google Scholar to identify evidence-based clinical practice guidelines pertaining to the use of PET, PET/computed tomography (CT), or PET/magnetic resonance in routine practice. We assessed the quality of each guideline using the Appraisal of Guidelines for Research and Evaluation II instrument and compared recommendations regarding indications for 18F-fluorodeoxyglucose (FDG) PET/CT. RESULTS: Thirty-five guidelines for PET imaging, published between 2008 and 2021, were included. These guidelines performed well in the domains of scope and purpose (median 80.6%, inter-quartile range [IQR] 77.8-83.3%) and clarity of presentation (median 75%, IQR 69.4-83.3%), but poorly in applicability (median 27.1%, IQR 22.9-37.5%). Recommendations for 48 indications in 13 cancers were compared. Considerable inconsistencies in the direction of whether to support the use of FDG PET/CT were observed in 10 (20.1%) indications pertaining to 8 cancer types: head and neck cancer (treatment response assessment), colorectal cancer (staging in patients with stages I-III disease), esophageal cancer (staging), breast cancer (restaging and treatment response assessment), cervical cancer (staging in patients with stage < IB2 disease and treatment response assessment), ovarian cancer (restaging), pancreatic cancer (diagnosis), and sarcoma (treatment response assessment). CONCLUSIONS: Current guidelines for PET imaging vary in methodological quality and provided considerably inconsistent recommendations. Efforts are needed to improve adherence to guideline development methodologies, to synthesis high-quality evidence, and to adopt standard terminologies. PROTOCOL REGISTRATION NUMBER: PROSPERO CRD42020184965. CLINICAL RELEVANCE STATEMENT: Guidelines for PET imaging provide considerably inconsistent recommendations and vary in methodological quality. It is suggested that clinicians be critical of these recommendations when applying them in practice, that guideline developers adopt more rigorous development methodologies, and that researchers prioritize research gaps identified by current guidelines. KEY POINTS: • PET guidelines vary in methodological quality and provided inconsistent recommendations. Efforts are needed to improve methodologies, synthesize high-quality evidence, and standardize terminologies. • Among six domains of methodological quality assessed by the AGREE II tool, guidelines for PET imaging performed well in scope and purpose (median 80.6%, inter-quartile range 77.8-83.3%) and clarity of presentation (75%, 69.4-83.3%), but poorly in applicability (27.1%, 22.9-37.5%). • Among the 48 recommendations (for 13 cancer types) compared, conflicts in the direction of whether to support FDG PET/CT use were observed in 10 (20.1%), for 8 cancer types (i.e., head and neck, colorectal, esophageal, breast, cervical, ovarian, pancreatic, and sarcoma).

Pulmonary Sclerosing Pneumocytoma on 18 F-FDG PET/MRI.

ABSTRACT: Pulmonary sclerosing pneumocytoma is a rare benign neoplasm. Owing to the low incidence, its radiographic features on 18 F-FDG PET/MRI are not well-known. Herein, we described findings of pulmonary sclerosing pneumocytoma on 18 F-FDG PET/MRI in a 52-year-old woman. It showed moderate FDG uptake and hyperintensity signal on both T1WI and T2WI images.

Study of quality of life and its correlated factors in patients after lumbar fusion for lumbar degenerative disc disease.

Background: In the present work, we aimed to explore the correlated factors of quality of life in patients receiving lumbar fusion for lumbar degenerative disc disease (DDD) in China. Methods: A total of 180 patients treated with lumbar fusion were included in the present study. Their general demographic characteristics, Visual Analog Scale (VAS) scores, Japanese Orthopedic Association (JOA) scores, Simplified Coping Style Questionnaire (SCSQ), Social Support Questionnaire (SSQ), and Medical Outcomes Study Short Form 36 (MOS SF-36) were collected and evaluated preoperatively and at 1 year postoperatively. Results: There were significant improvements in scores of VAS, JOA, and quality of life of patients from preoperation to 1-year postoperation after lumbar fusion. Marital status, with or without children, education level, economic pressure, and social support had significant predictive effects on the physical health of patients undergoing lumbar fusion. Marital status, education level, and economic pressure had significant predictive effects on the mental health of patients undergoing lumbar fusion. Conclusions: Factors correlated with the physical health of patients after lumbar fusion included positive coping style, negative coping style, social support, age, education level (high school college), disease duration (5-10), suffering from other diseases (combined with two or more other disease) and the number of surgical segments (double and three or more). Factors correlated with the mental health included negative coping style, social support, age, education level (middle school and high school college) and the number of surgical segments (double and three or more). The results verify that these factors were correlated to the patient's quality of life after lumbar fusion. Emphasizing and selectively intervening these correlated factors can further improve the quality of life in patients receiving lumbar fusion for lumbar degenerative disc disease.

Low-Grade Myofibroblastic Sarcoma Demonstrated on 99mTc-MDP Bone Scan and 18F-FDG PET/CT.

A 24-year-old woman with left hip pain for 4 months was admitted. Radiographs of the left hip showed a radiolucent lesion in the proximal femur. The lesion demonstrated hypointensity on T1-weighted and high intensity on T2-weighted fat saturated images. Tc-MDP bone scan and F-FDG PET/CT illustrated intense radiotracer accumulation at the osteolytic lesion. Postoperative pathology confirmed a low-grade myofibroblastic sarcoma (LGMS). LGMS is a rare neoplasm, which often occurred in the head and neck region. Our case suggested that LGMS should be considered in the differential diagnosis of proximal femur lesion on Tc-MDP bone scan and F-FDG PET/CT.

Current status and quality of radiomics studies in lymphoma: a systematic review.

OBJECTIVES: To perform a systematic review regarding the developments in the field of radiomics in lymphoma. To evaluate the quality of included articles by the Quality Assessment of Diagnostic Accuracy Studies-2 (QUADAS-2), the phases classification criteria for image mining studies, and the radiomics quality scoring (RQS) tool. METHODS: We searched for eligible articles in the MEDLINE/PubMed and EMBASE databases using the terms "radiomics", "texture" and "lymphoma". The included studies were divided into two categories: diagnosis-, therapy response- and outcome-related studies. The diagnosis-related studies were evaluated using the QUADAS-2; all studies were evaluated using the phases classification criteria for image mining studies and the RQS tool by two reviewers. RESULTS: Forty-five studies were included; thirteen papers (28.9%) focused on the differential diagnosis of primary central nervous system lymphoma (PCNSL) and glioblastoma (GBM). Thirty-two (71.1%) studies were classified as discovery science according to the phase classification criteria for image mining studies. The mean RQS score of all studies was 14.2% (ranging from 0.0 to 40.3%), and 23 studies (51.1%) were given a score of < 10%. CONCLUSION: The radiomics features could serve as diagnostic and prognostic indicators in lymphoma. However, the current conclusions should be interpreted with caution due to the suboptimal quality of the studies. In order to introduce radiomics into lymphoma clinical settings, the lesion segmentation and selection, the influence of the pathological pattern and the extraction of multiple modalities and multiple time points features need to be further studied. KEY POINTS: • The radiomics approach may provide useful information for diagnosis, prediction of the therapy response, and outcome of lymphoma. • The quality of published radiomics studies in lymphoma has been suboptimal to date. • More studies are needed to examine lesion selection and segmentation, the influence of pathological patterns, and the extraction of multiple modalities and multiple time point features.

Application of a modified surgical position in anterior approach for total cervical artificial disc replacement.

BACKGROUND: Total cervical artificial disc replacement (TDR) has been considered a safe and effective alternative surgical treatment for cervical spondylosis and degenerative disc disease that have failed to improve with conservative methods. Positioning the surgical patient is a critical part of the procedure. Appropriate patient positioning is crucial not only for the safety of the patient but also for optimizing surgical exposure, ensuring adequate and safe anesthesia, and allowing the surgeon to operate comfortably during lengthy procedures. The surgical posture is the traditional position used in anterior cervical approach; in general, patients are in a supine position with a pad under their shoulders and a ring-shaped pillow under their head. AIM: To investigate the clinical outcomes of the use of a modified surgical position versus the traditional surgical position in anterior approach for TDR. METHODS: In the modified position group, the patients had a soft pillow under their neck, and their jaw and both shoulders were fixed with wide tape. The analyzed data included intraoperative blood loss, position setting time, total operation time, and perioperative blood pressure and heart rate. RESULTS: Blood pressure and heart rate were not significantly different before and after body positioning in both groups (P > 0.05). Compared with the traditional position group, the modified position group showed a statistically significantly longer position setting time (P < 0.05). However, the total operation time and intraoperative blood loss were significantly reduced in the modified position group compared with the traditional position group (P < 0.05). CONCLUSION: The clinical outcomes indicated that total operation time and intraoperative blood loss were relatively lower in the modified position group than in the traditional position group, thus reducing the risks of surgery while increasing the position setting time. The modified surgical position is a safe and effective method to be used in anterior approach for TDR surgery.

A Nanoemulsion with A Porphyrin Shell for Cancer Theranostics.

A nanoemulsion with a porphyrin shell (NewPS) was created by the self-assembly of porphyrin salt around an oil core. The NewPS system has excellent colloidal stability, is amenable to different porphyrin salts and oils, and is capable of co-loading with chemotherapeutics. The porphyrin salt shell enables porphyrin-dependent optical tunability. The NewPS consisting of pyropheophorbide a mono-salt has a porphyrin shell of ordered J-aggregates, which produced a narrow, red-shifted Q-band with increased absorbance. Upon nanostructure dissociation, the fluorescence and photodynamic reactivity of the porphyrin monomers are restored. The spectrally distinct photoacoustic imaging (at 715 nm by intact NewPS) and fluorescence increase (at 671 nm by disrupted NewPS) allow the monitoring of NewPS accumulation and disruption in mice bearing KB tumors to guide effective photodynamic therapy. Substituting the oil core with Lipiodol affords additional CT contrast, whereas loading paclitaxel into NewPS facilitates drug delivery.

Endoscopic partial closure followed by adequate drainage for treating delayed perforation caused by duodenal endoscopic submucosal dissection: A case report.

RATIONALE: Delayed perforation of duodenal endoscopic submucosal dissection (ESD) was reported to be up to 14.3%. High invasive surgery remains the main treatment for delayed duodenal perforation. PATIENT CONCERNS: A 56-year-old woman presented with abdominal pain and fever at 1st day after ESD for treating a large laterally spreading tumor in the second part of duodenum. DIAGNOSIS: Emergent abdominal computed tomography revealed the presence of duodenal perforation. INTERVENTIONS: Endoscopic purse-string technique was used to partially close the large mucosal defect. Percutaneous endoscopic gastrostomy was conducted for gastric drainage and proximal drainage of the wound. A nasointestinal decompression tube was placed for distal drainage of the wound. OUTCOMES: No further symptoms were noted after 5 days. Both upper gastrointestinal series and endoscopy confirmed the healing of the wound. LESSONS: Partial closure of the mucosal defect followed by adequate drainage can be selected as a preferred choice for management of delayed duodenal perforation. It may also serve as an alternative for prevention of delayed perforation when complete closure of the mucosal defect is technically difficult or impossible.

Matrix metallopeptidase 2 targeted delivery of gold nanostars decorated with IR-780 iodide for dual-modal imaging and enhanced photothermal/photodynamic therapy.

Nanotheranostics has gained increasing interest, as it offers a great potential to realize personalized diagnostics and therapy. In this work, we report a facile approach of the fabrication of gold nanostars (GNS) attached with matrix metalloproteinases (MMP2) polypeptides (Ac-GPLGIAGQ) and IR-780 iodide through bovine serum albumin (BSA) for targeted dual-modal photoacoustic (PA)/near-infrared (NIR) fluorescence imaging and enhanced photothermal therapy (PTT)/photodynamic therapy (PDT) for lung cancer. MMP2 polypeptides served as the targeting ligand, IR-780 iodide functioned as the NIR fluorescence imaging agent as well as PTT/PDT agent, and GNS acted as the carrier of IR-780 molecules and performed PA imaging and PTT. DLS and CCK-8 assay demonstrated that the nanoprobes (GNS@BSA/I-MMP2) exhibited excellent stability and biocompatibility under physiological conditions. Subsequent in vitro studies verified that GNS@BSA/I-MMP2 nanoparticles (NPs) were effectively internalized by A549 cancer cells and exhibited remarkable antitumor efficacy. Furthermore, GNS@BSA/I-MMP2 NPs could specifically target the tumor and significantly suppress the tumor growth, and their antitumor effects were mainly through the synergistic effects of PDT and PTT based on IR-780 and GNS. These findings imply the potential of GNS@BSA/I-MMP2 NPs as a targeting PA/NIR probe in tumor diagnosis and combined therapy with a single light source. STATEMENT OF SIGNIFICANCE: We reported a convenient and facile approach to load IR-780 iodides in gold nanostars (GNS). This material could simultaneously perform near-infrared imaging/photoacoustic imaging and thermotherapy/photodynamic therapy. MMP2 coating on the surface of GNS@BSA/IR-780 promoted the prepared nanoparticles (GNS@BSA/I-MMP2) to target the tumor region. The heat generated by the synergistic effect of the GNS and IR-780 molecules resulted in the high temperature of the GNS@BSA/I-MMP2 NPs, which efficiently suppressed the growth of tumor, and the tumor volume decreased by 93% compared with that in the PBS groups with laser irradiation.

Gd3+-Ion-induced carbon-dots self-assembly aggregates loaded with a photosensitizer for enhanced fluorescence/MRI dual imaging and antitumor therapy.

The development of multifunctional nanoparticles for tumor theranostics has become a research hotspot. Despite the advantages of non-invasive precision diagnostics and efficient drug-delivery, these nanoparticles bring two significant issues: (i) a potential toxic effect and (ii) difficult clearance. To solve these issues, carbon dots (C-dots) are key potential candidates owing to their unique properties, such as excellent biocompatibility and rapid renal clearance. However, their small size leads to a short circulation time in the blood, which causes non-sufficient tumor accumulation for antitumor therapy. To reach the balance between an efficient accumulation in a tumor and rapid clearance from the body, herein we report a new multifunctional nanoprobe: photosensitizer (chlorine e6, Ce6)-loaded assembled C-dots (A-C-dots@Ce6). The A-C-dots@Ce6 were assembled from negatively-charged discrete C-dots using Gd3+ ions as a "glue". which also provided another function of in vivo nanoprobe monitoring via magnetic resonance (MR) imaging. Moreover, the nanoprobe exhibited an acidic pH-dependent disassembly and drug-release property. Benefiting from these advantages, the nanoprobe showed a targeted antitumor effect in A549 tumor-bearing mice under laser irradiation and gradual disassembly in the tumor for later body clearance. Therefore, the nanoprobe potentially provides a new strategy to solve the above balance issue, and brightens the future for antitumor monitoring and treatment.

Mimicking Pathogenic Invasion with the Complexes of Au22(SG)18-Engineered Assemblies and Folic Acid.

Biological systems provide the richest spectrum of sophisticated design for materials engineering. We herein provide a paradigm of Au22(SG)18-engineered (SG, glutathione thiolate) and hydrogen bonds engaged assemblies for mimicking capsid protein self-assembly. The water-evaporation-induced self-assembly method allows discrete ultrasmall gold nanoclusters (GNCs) to be self-assembled into super-GNCs assemblies (SGNCs) ranging from nano-, meso- to microscale in water-dimethyl sulfoxide binary solvents in a template-free manner. After removing free and hydration layer water molecules, the formation of SGNCs is engaged by the collective cohesion of hydrogen bonds between glutathione ligands of gradually approaching GNCs. Then, a series of tightly orchestrated cellular events induced by the complexes of Au22(SG)18-engineered assemblies and folic acid are demonstrated to mimic the invasion of eukaryotic cells by pathogens. First, the activation of macropinocytosis mimics the macropinocytic entry used by the pathogens to invade host cells. Then the cytoplasmic vacuolization is a mimicry of vacuolating effects induced by the oligomeric vacuolating toxins secreted by some bacteria. Lastly, the escaping from macropinosomes into cytosol is in a vacuolating toxin's strategy. The findings demonstrate the capabilities of artificial pathogens to emulate the structures and functions of natural pathogens.

Cytokine induced killer cells-assisted delivery of chlorin e6 mediated self-assembled gold nanoclusters to tumors for imaging and immuno-photodynamic therapy.

The cytotoxicity and unique tumor-tropic properties of cytokine-induced killer (CIK) cells render them promising in the field of cancer immunotherapy and delivery systems. Here, we report a novel and facile approach to assemble gold nanoclusters (GNCs) into stable and monodispersed nanoparticles (NPs) using Chlorin e6 (Ce6) molecules. Notably, the fluorescence intensity of the GNCs-Ce6 NPs was about 4.5 folds stronger than the GNCs counterparts. The as-prepared GNCs-Ce6 NPs were conjugated with CD3 antibody (Ab) and further employed to label CIK cells to create a CIK cell-based drug delivery system (Ce6-GNCs-Ab-CIK). The Ce6-GNCs-Ab-CIK exhibited high tumor-targeting efficiency and excellent therapeutic efficacy toward MGC-803 tumor-bearing mice. Benefiting from the synergistic therapeutic effect between GNCs-Ce6-Ab NPs and CIK cells, the GNCs-Ce6-Ab-CIK strategy may present an ideal cancer theranostic platform for tumor targeted imaging and combination therapy.

pH-responsive gold nanoclusters-based nanoprobes for lung cancer targeted near-infrared fluorescence imaging and chemo-photodynamic therapy.

Nanoparticle-based drug delivery systems have drawn a great deal of attention for their opportunities to improve cancer treatments over intrinsic limits of conventional cancer therapies. Herein, we developed the polypeptide-modified gold nanoclusters (GNCs)-based nanoprobes for tumor-targeted near-infrared fluorescence imaging and chemo-photodynamic therapy. The nanoprobes comprise of tetra-functional components: i) polyethylene glycol (PEG) shell for long blood circulation and better biocompatibility; ii) MMP2 polypeptide (CPLGVRGRGDS) for tumor targeting; iii) cis-aconitic anhydride-modified doxorubicin (CAD) for pH-sensitive drug release; iv) photosensitizer chlorin e6 (Ce6) for photodynamic therapy and fluorescence imaging. The in vitro results demonstrated that the as-synthesized nanoprobes could be efficiently internalized into A549 cells and then significantly enhance the mortality of cancer cells compared with free Ce6 and doxorubicin. For in vivo tests, the nanoprobes showed excellent tumor targeting ability, long blood circulation time, and could remarkably inhibit the growth of tumor. Our results will help to advance the design of combination strategies to enhance the efficacy of imaging-guided cancer therapy. STATEMENT OF SIGNIFICANCE: The as-prepared CDGM NPs could accumulate into the tumor tissue with the enhanced permeability and retention (EPR) effect as well as the active tumor targeting ability from the MMP2 polypeptides. With the acid-sensitive linker, the doxorubicin (DOX) would be released from the synthesized nanoparticles after exposing to the acid tumor microenvironment. The CDGM NPs exhibit excellent tumor targeting ability and could remarkably suppress the growth of tumor compared with free Ce6 and DOX.

Two-phase electrospinning to incorporate growth factors loaded chitosan nanoparticles into electrospun fibrous scaffolds for bioactivity retention and cartilage regeneration.

Growth factor is an essential ingredient to regulate mesenchymal stem cells (MSCs) chondrogenic differentiation in cartilage tissue engineering. However, non-osteochondral specification, short plasma half time and bioactivity loss restrict growth factor's application. Thus, novel chondrogenic growth factors, specifically target osteochondral lineage cells, that can be sustained release and bioactivity protected to exert functions continually and effectively have attracted increasing researchers' interest. To achieve these goals, chitosan nanoparticles and electrospun fiber scaffolds were used as dual release system to sustain release Nel-like molecule-1 (Nell-1) growth factor and protect bioactivity, then the effect and mechanism of Nell-1 on inducing human bone MSCs (hBMSCs) differentiate toward chondrocytes were investigated. For release and bioactivity protection study, preloading Nell-1 into chitosan nanoparticles significantly extended the release time, increased the released Nell-1's bioactivity than directly incorporating Nell-1 into the scaffolds. Furthermore, Nell-1 specifically promotes hBMSCs in vitro chondrogenic differentiation by increasing expression of chondrogenic related genes and proteins. These findings suggest the potential utility of Nell-1 incorporated dual release scaffold for cartilage tissue engineering.

Tumor-triggered drug release from calcium carbonate-encapsulated gold nanostars for near-infrared photodynamic/photothermal combination antitumor therapy.

Different stimulus including pH, light and temperature have been used for controlled drug release to prevent drug inactivation and minimize side-effects. Herein a novel nano-platform (GNS@CaCO3/ICG) consisting of calcium carbonate-encapsulated gold nanostars loaded with ICG was established to couple the photothermal properties of gold nanostars (GNSs) and the photodynamic properties of indocyanine green (ICG) in the photodynamic/photothermal combination therapy (PDT/PTT). In this study, the calcium carbonate worked not only a drug keeper to entrap ICG on the surface of GNSs in the form of a stable aggregate which was protected from blood clearance, but also as the a pH-responder to achieve highly effective tumor-triggered drug release locally. The application of GNS@CaCO3/ICG for in vitro and in vivo therapy achieved the combined antitumor effects upon the NIR irradiation, which was superior to the single PDT or PTT. Meanwhile, the distinct pH-triggered drug release performance of GNS@CaCO3/ICG implemented the tumor-targeted NIR fluorescence imaging. In addition, we monitored the bio-distribution and excretion pathway of GNS@CaCO3/ICG based on the NIR fluorescence from ICG and two-photon fluorescence and photoacoustic signal from GNSs, and the results proved that GNS@CaCO3/ICG had a great ability for tumor-specific and tumor-triggered drug release. We therefore conclude that the GNS@CaCO3/ICG holds great promise for clinical applications in anti-tumor therapy with tumor imaging or drug tracing.