Protein-nanoparticle co-assembly supraparticles for drug delivery: Ultrahigh drug loading and colloidal stability, and instant and complete lysosomal drug release.

Two frequent problems hindering clinical translation of nanomedicine are low drug loading and low colloidal stability. Previous efforts to achieve ultrahigh drug loading (>30 %) introduce new hurdles, including lower colloidal stability and others, for clinical translation. Herein, we report a new class of drug nano-carriers based on our recent finding in protein-nanoparticle co-assembly supraparticle (PNCAS), with both ultrahigh drug loading (58 % for doxorubicin, i.e., DOX) and ultrahigh colloidal stability (no significant change in hydrodynamic size after one year). We further show that our PNCAS-based drug nano-carrier possesses a built-in environment-responsive drug release feature: once in lysosomes, the loaded drug molecules are released instantly (<1 min) and completely (∼100 %). Our PNCAS-based drug delivery system is spontaneously formed by simple mixing of hydrophobic nanoparticles, albumin and drugs. Several issues related to industrial production are studied. The ultrahigh drug loading and stability of DOX-loaded PNCAS enabled the delivery of an exceptionally high dose of DOX into a mouse model of breast cancer, yielding high efficacy and no observed toxicity. With further developments, our PNCAS-based delivery systems could serve as a platform technology to meet the multiple requirements of clinical translation of nanomedicines.

Universal primer multiplex PCR assay for detection and genotyping of porcine astroviruses.

Porcine astroviruses (PAstV) are members of the family Astroviridae, Mamastravirus genus and have been identified to have five genotypes (PAstV1-5). These viruses are highly prevalent in pigs and can cause enteric disease as well as neurological or respiratory symptoms depending on their genotypes. At present, the epidemiological impacts of some PAstV genotypes on pigs are largely unknown and hence continuously monitoring of these PAstVs may be needed. The purpose of this research was to develop an improved and efficient detection tool for PAstVs and to evaluate the developed method using clinical samples. Initially, a set of five chimeric primers (CP), each comprising genotype specific primer pairs with an identical universal adapter at the 5' end, and a universal primer (UP) that is identical to universal adapter sequence, were designed. With these tools in place, a novel multiplex PCR system with universal primer was established for the simultaneous detection of the five types of PAstV. This method can specifically detect PAstV genotypes, with a limit of detection (LOD) of 5 copies/µL for each genotype irrespective of single or mixed target template. Using this new assay, 273 pig fecal samples were investigated for further assay evaluation. Among all samples, the positive rate was 70.0% with PAstV4 in 56.8% of the samples, PAstV2 in 38.8%, PAstV1 in 16.8%, and PAstV5 in 11.0%. More than one PAstV in a sample were detected in 39.2% of the samples. The consistency rate between the novel multiplex PCR and singleplex PCRs was 96.4-100%. Given its rapidity, specificity and sensitivity, the novel multiplex PCR is a useful approach for demonstrating single or mixed genotype infections of PAstV.

Biosynthesis and Gene Regulation of Rhamnolipid Congeners.

Rhamnolipid congeners have been widely used in agriculture and biomedicine as potent surfactants. They have recently attracted attention due to their diverse and versatile biological functions, which include an important bacterial virulence factor that makes them attractive targets for research into biosynthetic pathways and gene regulation. The intricate gene expression and regulation network controlling their biosynthesis remain to be completely understood. This article summarizes current knowledge about the biosynthesis pathways and regulatory mechanisms of rhamnolipid congeners, that meet the pharmacological needs of human health and agriculture.

Nanoplatforms Potentiated Ablation-Immune Synergistic Therapy through Improving Local Control and Suppressing Recurrent Metastasis.

Minimally invasive ablation has been widely applied for treatment of various solid tumors, including hepatocellular carcinoma, renal cell carcinoma, breast carcinomas, etc. In addition to removing the primary tumor lesion, ablative techniques are also capable of improving the anti-tumor immune response by inducing immunogenic tumor cell death and modulating the tumor immune microenvironment, which may be of great benefit to inhibit the recurrent metastasis of residual tumor. However, the short-acting activated anti-tumor immunity of post-ablation will rapidly reverse into an immunosuppressive state, and the recurrent metastasis owing to incomplete ablation is closely associated with a dismal prognosis for the patients. In recent years, numerous nanoplatforms have been developed to improve the local ablative effect through enhancing the targeting delivery and combining it with chemotherapy. Particularly, amplifying the anti-tumor immune stimulus signal, modulating the immunosuppressive microenvironment, and improving the anti-tumor immune response with the versatile nanoplatforms have heralded great application prospects for improving the local control and preventing tumor recurrence and distant metastasis. This review discusses recent advances in nanoplatform-potentiated ablation-immune synergistic tumor therapy, focusing on common ablation techniques including radiofrequency, microwave, laser, and high-intensity focused ultrasound ablation, cryoablation, and magnetic hyperthermia ablation, etc. We discuss the advantages and challenges of the corresponding therapies and propose possible directions for future research, which is expected to provide references for improving the traditional ablation efficacy.

Selective poly adenylation predicts the efficacy of immunotherapy in patients with lung adenocarcinoma by multiple omics research.

The aim of this study was to find the application value of selective polyadenylation in immune cell infiltration, biological transcription function and risk assessment of survival and prognosis in lung adenocarcinoma (LUAD). The processed original mRNA expression data of LUAD were downloaded, and the expression profiles of 594 patient samples were collected. The (APA) events in TCGA-NA-SEQ data were evaluated by polyadenylation site use Index (PDUI) values, and the invasion of stromal cells and immune cells and tumor purity were calculated to group and select the differential genes. Lasso regression and stratified analysis were used to examine the role of risk scores in predicting patient outcomes. The study also used the GDSC database to predict the chemotherapeutic sensitivity of each tumor sample and used a regression method to obtain an IC50 estimate for each specific chemotherapeutic drug treatment. Then CIBERSORT algorithm was used to conduct Spearman correlation analysis, immune regulatory factor analysis and TIDE immune system function analysis for gene expression level and immune cell content. Finally, the Kaplan-Meier curve was used to analyze the correlation between stromal score and the immune score of LUAD. In this study, APA's LUAD risk score prognostic model was constructed. KM survival analysis showed that immune score affected the prognosis of LUAD patients ( P = 0.027) but the matrix score was not statistically significant ( P = 0.1). We extracted 108 genes with APA events from 827 different genes and based on PUDI clustering and heat map, the survival rate of patients in the four groups was significantly different ( P = 0.05). Multiple omics studies showed that risk score was significantly positively correlated with Macrophages M0, T cells Follicular helper, B cells naive and NK cells resting. It is significantly negatively correlated with dendritic cells resting, mast cells resting, monocyte, T cells CD4 memory resting and B cells memory. We further explored the relationship between the expression of immunosuppressor genes and risk score and found that ADORA2A, BTLA, CD160, CD244, CD274, CD96, CSF1R and CTLA4 genes were highly correlated with the risk score. Selective poly adenylation plays an important role in the development and progression of LUAD, immune invasion, tumor cell invasion and metastasis and biological transcription, and affects the survival and prognosis of LUAD patients.

Soil Respiration of Paddy Soils Were Stimulated by Semiconductor Minerals.

Large quantities of semiconductor minerals on soil surfaces have a sensitive photoelectric response. These semiconductor minerals generate photo-electrons and photo-hole pairs that can stimulate soil oxidation-reduction reactions when exposed to sunlight. We speculated that the photocatalysis of semiconductor minerals would affect soil carbon cycles. As the main component of the carbon cycle, soil respiration from paddy soil is often ignored. Five rice cropping areas in China were chosen for soil sampling. Semiconductor minerals were measured, and three main semiconductor minerals including hematile, rutile, and manganosite were identified in the paddy soils. The identified semiconductor minerals consisted of iron, manganese, and titanium oxides. Content of Fe2O3, TiO2, and MnO in the sampled soil was between 4.21-14%, 0.91-2.72%, and 0.02-0.22%, respectively. Most abundant semiconductor mineral was found in the DBDJ rice cropping area in Jilin province, with the highest content of Fe2O3 of 14%. Soils from the five main rice cropping areas were also identified as having strong photoelectric response characteristics. The highest photoelectric response was found in the DBDJ rice cropping area in Jilin province with a maximum photocurrent density of 0.48 µA/cm2. Soil respiration was monitored under both dark and light (3,000 lux light density) conditions. Soil respiration rates in the five regions were (from highest to lowest): DBDJ > XNDJ > XBDJ > HZSJ > HNSJ. Soil respiration was positively correlated with semiconductor mineral content, and soil respiration was higher under the light treatment than the dark treatment in every rice cropping area. This result suggested that soil respiration was stimulated by semiconductor mineral photocatalysis. This analysis provided indirect evidence of the effect semiconductor mineral photocatalysis has on the carbon cycle within paddy soils, while exploring carbon conversion mechanisms that could provide a new perspective on the soil carbon cycle.

Rare soil bacteria are more responsive in desertification restoration than abundant bacteria.

Soil microbes play key roles in ecosystem functions, especially in the recovery of ecosystems from disturbance, and exploring community assembly under changing environments has long been a central theme in microbial ecology. The response of abundant and rare bacteria in desertified land to restoration is still unclear. Here, we investigated the effects of vegetation restoration on the assemblage patterns of abundant and rare bacteria in soil across the four sandy lands (Hulunbeir, Horqin, Otindag, and Mu Us) in northern China. Our results revealed that abundant bacteria maintained a relatively stable state under restoration, whereas rare taxa were more responsive, indicating the higher resilience of the rare community to change. Our network analysis also showed that restoration promoted destabilizing properties in rare, but not in abundant, bacterial co-occurrence networks in soil. Environmental selection played a key role in abundant and rare community assembly under restoration. Of the two, the rare subcommunity was mainly affected by environmental filtering. The variations in the abundant and rare communities at the sampling sites under restoration were controlled mainly by plant species richness, and stronger effects were observed in the rare taxa. Overall, these results provide new insight into the mechanisms controlling bacterial community assembly in response to vegetation restoration.

[Effects of cigarette smoke extract on mitochondrial function of macrophages].

Objective: To investigate the effects of cigarette smoke extract (CSE) on the mitochondrial function of macrophages. Methods: RAW264.7 macrophages were used for the experiment in this study. When the cell density was about 70%, the old culture medium was abandoned, and the 100% CSE stock solution was diluted with serum-free DMEM and FBS into 1%, 5%, 15%, 25% and 90% CSE and added to the well plate. The cell activity of RAW264.7 cells treated with CSE at different concentrations for 24 h was detected by CCK-8 method. Then the optimal CSE concentration was selected to treat cells for 0 h, 24 h, 48 h or 72 h respectively, and CCK-8 assay was used to detect the cell activity of CSE treated cells at different time groups. After the cells were treated with 0%, 5% and 25% CSE for 24 hours, cell necrosis and apoptosis was detected by Annexin V-FITC /PI staining; Mitochondrial membrane damage of RAW 264.7 was detected by mitochondrial membrane potential assay kit with JC-1; Macrophages were stained with ROS-specific dye DCFH-DA, and then Flow cytometer was used to determine the fluorescence and the proportion of ROS-positive macrophages; the enhanced ATP assay kit was used to detect the intracellular ATP concentration. Results: ①Compared with 0% CSE, cell viability was increased significantly in 1% CSE group (P＜0.01), cell viability was decreased significantly when CSE concentration was above 5% (P＜0.05); Macrophages were treated with 5% CSE, and cell viability was decreased significantly with the increase of treatment time (P＜0.01). ②Compared with 0% CSE, 5% CSE and 25% CSE mainly caused macrophage necrosis, decreased mitochondrial membrane potential, increased ROS production and decreased ATP significantly (P＜0.05 or P＜0.01), and the changes were more significant in 25% CSE treatment group(P＜0.05 or P＜0.01). Conclusion: CSE may affect mitochondrial function of macrophages, leading to decreased cell viability and necrosis.

Novel universal primer-pentaplex PCR assay based on chimeric primers for simultaneous detection of five common pig viruses associated with diarrhea.

Viral pathogens associated with diarrhea in pigs include porcine circovirus 2 (PCV2), porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV), porcine rotavirus A (RVA) and C (RVC) among others. In this study, a novel universal primer-based pentaplex PCR (UP-M-PCR) assay was developed for simultaneous detection and differentiation of these five viruses. The assay uses a short-cycle multiplex amplification by chimeric primers (CP), which are virus specific, with a tail added at the 5' end of the universal primer (UP), followed by universal amplification using UPs and a regular cycle amplification. Five universal primers with CPs (UP1-5) were designed and evaluated in an UP-based single PCR (UP-S-PCR). All five UPs were found to work efficiently and UP2 exhibited the best performance. After system optimizations, the analytical sensitivity of the UP-M-PCR, using plasmids containing the specific viral target fragments, was 5 copies/reaction for each of the five viruses irrespective of presence of a single or multiple viruses in the reaction. No cross-reaction was observed with other non-target viruses. When 273 fecal samples from clinically healthy pigs were tested, the assay sensitivity was 90.9-100%, the specificity was 98.0-100%, and the agreement rate with the UP-S-PCR was 98.5-99.6% with a Kappa value being 0.95-0.98. In summary, the UP-M-PCR developed here is a rapid and highly sensitive and specific detection method that can be used to demonstrate mixed infections in pigs with diarrhea.

Immunogenicity and safety of a live attenuated varicella vaccine in children 1-12 years of age: A randomized, blinded, controlled, non-inferiority phase 3 clinical trial.

OBJECTIVES: To evaluate the immunogenicity and safety of a live attenuated varicella vaccine produced using a cell factory process. METHODS: In this randomized, blinded, controlled, non-inferiority phase 3 clinical trial conducted in Guizhou, healthy children aged 1-12 years were randomly assigned in a 2: 1 ratio to receive one dose of experimental or control vaccine. Physical examination and first blood collection were performed preimmunization on day 0. Diary cards were collected after day 15. Contact cards and second blood samples were collected on day 30. The primary immunogenicity endpoint was the positive conversion rate of the anti-varicella virus antibody at 30 days postimmunization in susceptible children. Secondary endpoints were the fourfold increase rate, positive conversion rate, geometric mean titer, and geometric mean increase at 30 days after immunization in the total cohort. RESULTS: Of the 900 children assessed for eligibility, 894 received an experimental or control vaccine. Both the full analysis and safety analysis sets included 894 subjects. The seroconversion rate in the susceptible population was 95.84% in the experimental and 94.76% in the control group. The lower limit of the 95% confidence interval difference was -2.37%, which was greater than the non-inferiority margin set by the program (-10%). No significant difference in solicited adverse reactions was found between the groups. Within 6 months postimmunization, a total of 24 serious adverse events were reported, none related to the studied vaccine. CONCLUSION: The live attenuated varicella vaccine produced using a cell factory process was highly immunogenic, safe, and non-inferior to the product in the market. Further studies need to be implemented in the immune persistence, the epidemiological effectiveness and the rare adverse reactions.

Chaetomugilin J Enhances Apoptosis in Human Ovarian Cancer A2780 Cells Induced by Cisplatin Through Inhibiting Pink1/Parkin Mediated Mitophagy.

PURPOSE: The chemoresistance and toxicity of traditional chemotherapeutic drugs have become obstacles to their antitumor effects in ovarian cancers. Therefore, it is particularly important to develop new anticancer drugs to increase target sensitivity and reduce the toxicity of chemotherapy drugs. As key organelles, the endoplasmic reticulum and mitochondria play important role in chemoresistance. Cells become resistant to drugs by maintaining the homeostasis of the endoplasmic reticulum and mitochondria. Chaetomugilin J, a metabolite isolated from Polygonatum sibiricum, belongs to the Chaetomium family and exhibits potent cytotoxicity. In this study, we aimed to explore the mechanistic link between apoptosis and endoplasmic reticulum stress, mitophagy and mitochondrial dysfunction induced by chaetomugilin J combined with cisplatin in the ovarian cancer cell line A2780. METHODS: Chaetomugilin J was identified by chemical methods. Cell viability was measured by an MTT assay. The apoptosis, mitochondrial membrane potential, and intracellular reactive oxygen species (ROS) were examined by flow cytometry. Mitochondrial ROS was measured by a fluorescence microscope with MitoSox staining. Further, the related proteins and overexpression of parkin were detected by Western blot. RESULTS: Chaetomugilin J combined with low-dose cisplatin decreased cell viability and increased apoptosis in A2780 cells. In addition, intracellular ROS and mitochondrial ROS were increased, while the mitochondrial membrane potential was reduced. The expressions of grp78 and chop were decreased after treatment by chaetomugilin J combined with low-dose cisplatin. Overexpression of parkin attenuated chaetomugilin J combined with cisplatin-induced apoptosis. CONCLUSION: Chaetomugilin J combined with cisplatin inhibited pink1/parkin mediated mitophagy increased mitochondrial dysfunction in the A2780 cells and enhanced apoptosis induced by cisplatin in the ovarian cancer cell line A2780. But this process was not related to endoplasmic reticulum apoptotic pathway.

Portable Handheld Slit-Lamp Based on a Smartphone Camera for Cataract Screening.

PURPOSE: As part of plans to provide help to people in remote and poor areas who have no medical resources, a portable slit-lamp based on a smartphone was proposed. This would help in early screening of cataract diseases. METHODS: This means a microlens is designed that would work with a phone's camera. The phone's photo taking function is used in capturing the image of the eyes lens to replace the observation system of the desktop slit-lamp. A simplified slit light band was designed. In order for the light source part to meet the portable requirements of the slit-lamp, the adjustable and diffused light functions of the ligaments were removed in this design. Furthermore, the images collected by the smartphone are uploaded to the deep learning cataract screening system, which can achieve real-time and effective screening of cataract. RESULTS: Unlike the desktop slit-lamp, which needs skilled personnel to operate, this device can be easily operated by less-skilled or inexperienced doctors. This eliminates the concerns of inaccurate diagnosis based on the use of unskilled professionals. Due to the portability, ease of use, and simplicity in obtaining crystal images of this device, it serves as a promising platform for nonhospital screening and telemedicine. CONCLUSIONS: In this paper, we invented a small portable device for screening cataract. This device is to make screening and diagnosis of cataract in remote areas very fast and effective. It will also solve the problem of inadequate specialized doctors and equipment in those areas as well. Translational Relevance. Smartphones can be used with portable slit-lamps to capture the images of the lens.

Hydrochromic CsPbBr3 Nanocrystals for Anti-Counterfeiting.

Hydrochromic materials that can reversibly change color upon water treatment have attracted much attention owing to their potential applications in diverse fields. Herein, for the first time, we report that space-confined CsPbBr3 nanocrystals (NCs) are hydrochromic. When CsPbBr3 NCs are loaded into a porous matrix, reversible transition between luminescent CsPbBr3 and non-luminescent CsPb2 Br5 can be achieved upon the exposure/removal of water. The potential applications of hydrochromic CsPbBr3 NCs in anti-counterfeiting are demonstrated by using CsPbBr3 NCs@mesoporous silica nanospheres (around 100 nm) as the starting material. Owing to the small particle size and negatively charged surface, the as-prepared particles can be laser-jet printed with high precision and high speed. We demonstrate the excellent stability over repeated transformation cycles without color fade. This new discovery may not only deepen the understanding of CsPbX3 , but also open a new way to design CsPbX3 materials for new applications.

Producing protein-nanoparticle co-assembly supraparticles by the interfacial instability process.

Originally discovered in fundamental research of nanomaterial-biomolecule interactions, protein-nanoparticle co-assembly supraparticles (PNCAS) have become an emerging class of nanomaterials with various biological applications. We apply the interfacial instability process, which was originally reported for forming nanoparticles-encapsulated polymeric micelles, to produce PNCAS. By doing so hydrophobic nanoparticles, which are often the product formed from the upstream nanoparticle synthesis step, can be directly used as the raw materials of the production process of PNCAS. On the other hand, we take advantage of the structural features of protein molecules, in comparison with amphiphilic block copolymers, to mitigate two common problems encountered in the original interfacial instability-mediated nanoparticle encapsulation process, namely (1) poor encapsulation number control and (2) inconvenience and high cost to vary the assembly size. Additionally, we achieve semi-continuous and scalable production of PNCAS by combining the electrospray process and the interfacial instability process. We also conduct proof-of-concept studies of biological applications of the PNCAS products.

Research on Total Factor Productivity and Influential Factors of the Regional Water-Energy-Food Nexus: A Case Study on Inner Mongolia, China.

With the supply of water, energy and food facing severe challenges, there has been an increased recognition of the importance of studying the regional water-energy-food nexus. In this paper, Inner Mongolia, including 12 cities in China, was selected as a research case. A super-efficiency slack based measure (SBM) model that considered the undesirable outputs was adopted to calculate the regional total factor productivity (TFP) and the Malmquist-Luenberger index was used to investigate the change trend of the TFP from 2007 to 2016 based on understanding the water-energy-food nexus. Finally, influential factors of the TFP were explored by Tobit regression. The results show that the 12 Inner Mongolia cities are divided into higher, moderate and lower efficiency zones. The higher efficiency zone includes Ordos, Hohhot, Xing'an, and Tongliao, and the lower efficiency zone includes Chifeng, Xilin Gol, Baynnur, Wuhai and Alxa. There is a serious difference in TFP between Inner Mongolia cities. During the study period, the TFP of the water-energy-food nexus in Inner Mongolia cities shows a rising trend, which is mainly driven by the growth of technical progress change. However, the average ML values of the lower and moderate efficiency zones were inferior to the higher efficiency zone in six of the ten years, so the difference between Inner Mongolia cities is growing. According to the Tobit regression, the mechanization level and degree of opening up have positive effects on the TFP, while enterprise scale and the output of the third industry have negative effects on the TFP. Government support does not have any significant impact on the TFP. Finally, suggestions were put forward to improve the TFP of the water-energy-food nexus in Inner Mongolia cities.

Enhancing the effects of transcranial magnetic stimulation with intravenously injected magnetic nanoparticles.

Transcranial magnetic stimulation (TMS) is a non-invasive and clinically approved method for treating neurological disorders. However, the relatively weak intracranial electric current induced by TMS is an obvious inferiority which can only produce limited treatment effects in clinical application. The present study aimed to investigate the possibility of enhancing the effects of TMS with intravenously administrated magnetic nanoparticles. To facilitate crossing of the blood-brain barrier (BBB), the superparamagnetic iron oxide nanoparticles (SPIONs) were coated with carboxylated chitosan and poly(ethylene glycol). To aid the nanoparticles in crossing the BBB and targeting the predesigned brain regions, an external permanent magnet was attached to the foreheads of the rats before the intravenous administration of SPIONs. The electrophysiological tests showed that the maximum MEP amplitude recorded in an individual rat was significantly higher in the SPIONs + magnet group than in the saline group (5.78 ± 2.54 vs. 1.80 ± 1.55 mV, P = 0.015). In the M1 region, biochemical tests detected that the number density of c-fos positive cells in the SPIONs + magnet group was 3.44 fold that of the saline group. These results suggest that intravenously injected SPIONs can enhance the effects of TMS in treating neurological disorders.

Fabricating CsPbX3-Based Type I and Type II Heterostructures by Tuning the Halide Composition of Janus CsPbX3/ZrO2 Nanocrystals.

Fabricating CsPbX3-based heterostructures has proven to be a feasible way to tune their photophysical properties. Here, we report the successful fabrication of Janus CsPbX3/ZrO2 heterostructure nanocrystals (NCs), in which each CsPbX3 NC is partially covered by ZrO2. According to the band alignment, CsPbBr3/ZrO2 and CsPbI3/ZrO2 can be indexed as type I and type II composites, respectively. The type I composites display great enhancement in photoluminescence quantum yield (from 63 to 90%) and photoluminescence lifetime (from 12.9 to 66.1 ns) because of the charge carrier confinement and passivation effect provided by ZrO2. In contrast, the type II composites can be used in photocatalytic reduction of CO2 because electrons and holes are effectively separated and accumulated in ZrO2 and CsPbI3, respectively, under irradiation. Janus CsPbBr3/ZrO2 NCs showed a stability much higher than that of pristine CsPbBr3 against polar solvent treatment. A stable and highly efficient light-emitting device with luminous efficiency up to 55 lm W-1 is fabricated by using CsPbBr3/ZrO2 NCs as the green light source. This work may not only enrich the family of surface-passivated perovskite materials but also provide a good example for the rational design of specific composites in the metal halide perovskite field.

Intracellular targeted delivery of quantum dots with extraordinary performance enabled by a novel nanomaterial design.

Quantum dots (QDs) have emerged as a major class of fluorescent probes with unique optical properties, but applying QDs for imaging specific intracellular entities in live cells has been hindered by the poor performance of targeted intracellular delivery of QDs due to various cellular transport barriers. We describe a novel QD nanoprobe design, which is termed a cosolvent-bare hydrophobic QD-biomolecule (cS-bQD-BM, or 'SDot' for short), combining a cosolvent, a bare hydrophobic nanoparticle surface, ultrasmall size and biomolecular function. SDots show extraordinary intracellular targeting performance with the nucleus as the model target, including near-perfect specificity, excellent efficiency and reproducibility, high-throughput ability, minimal toxicity, and ease of operation, as well as superb optical properties and colloidal stability. We introduce integrated single-particle tracking and pair-correlation function analysis of a spinning-disk confocal microscope platform (iSPT-pCF-SDCM) to study SDot's cellular transport. Endocytosed SDots can undergo a highly potent and noninvasive process of vesicle escape, yielding complete vesicle escape with no serious vesicle disruption. We exploit SDots' unprecedented ability to overcome cellular transport barriers to enhance drug and macromolecule delivery.

Spontaneous and instant formation of highly stable protein-nanoparticle supraparticle co-assemblies driven by hydrophobic interaction.

Recently, supraparticle protein-nanoparticle co-assemblies (or 'supraparticle co-assemblies' for short) have attracted considerable interest due to their fundamental and technological value. However, it remains challenging to form supraparticle co-assemblies with high stability. Here, we show that using hydrophobic interaction, instead of the previously used electrostatic and van der Waals interactions, as the primary driving force can lead to instant formation of exceptionally stable supraparticle co-assemblies with minimal external energy input. Our formation method of supraparticle co-assemblies simply involves mixing globular proteins (e.g., bovine serum albumin) with hydrophobic nanoparticles (e.g., hydrophobic magnetic nanoparticles and hydrophobic quantum dots) without significant energy input (e.g., sonication or stirring). Upon mixing of hydrophobic nanoparticles and proteins, the formation of supraparticle co-assemblies only takes <1 minute. Further incubation of the mixture for several hours results in a gradual increase of the size uniformity of supraparticle co-assemblies. The formed supraparticle co-assemblies have been colloidally stable for 6 months and counting, and can withstand harsh environments such as basic and acidic pH, high temperature, high dilution, and serum. Co-encapsulation of different sizes/types of nanoparticles is found to be feasible and the co-encapsulation number ratio of different nanoparticles is well-controlled by the feeding ratio. Proof-of-concept studies show the potential of the supraparticle co-assemblies for biological imaging, delivery, and modulation. The combination of very rapid formation, minimal energy consumption, highly stable products, and inexpensive raw materials of this hydrophobic interaction-driven process meets many of the main goals of 'ideal' nano-manufacturing. Thus, this process could serve as the foundation of ideal manufacturing of supraparticle co-assemblies.

A potent, minimally invasive and simple strategy of enhancing intracellular targeted delivery of Tat peptide-conjugated quantum dots: organic solvent-based permeation enhancer.

Targeted delivery of nanomaterials to specific intracellular locations is essential for the development of many nanomaterials-based biological applications. Thus far the targeting performance has been limited due to various intracellular transport barriers, especially intracellular vesicle trapping. Here we report the application of permeation enhancers based on organic solvents in small percentage to enhance the intracellular targeted delivery of nanomaterials. Previously permeation enhancers based on organic solvents and ionic liquids have been used in overcoming biological transport barriers at tissue, organ, and cellular levels, but this strategy has so far rarely been examined for its potential in facilitating transport of nanometer-scale entities across intracellular barriers, particularly intracellular vesicle trapping. Using the cell nucleus as a model intracellular target and Tat peptide-conjugated quantum dots (QDs-Tat) as a model nanomaterial-based probe, we demonstrate that a small percentage (e.g. 1%) of organic solvent greatly enhances nucleus targeting specificity as well as increasing endocytosis-based cellular uptake of QDs. We combine vesicle colocalization (DiO dye staining), vesicle integrity (calcein dye release), and single-particle studies (pair-correlation function microscopy) to investigate the process of organic solvent-enhanced vesicle escape of QDs-Tat. The organic solvent based vesicle escape-enhancing approach is found to be not only very effective but minimally invasive, resulting in high vesicle escape efficiency with no significant disruption to the membrane integrity of either intracellular vesicles or cells. This approach drastically outperforms the commonly used vesicle escape-enhancing agent (i.e., chloroquine, whose enhancement effect is based on disrupting vesicle integrity) in both potency and minimal invasiveness. Finally, we apply organic solvent-based targeting enhancement to improve the intracellular delivery of the anticancer drug doxorubicin (DOX).