Particle Integrity and Size Effect on the Journey of Polymeric Nanocarriers in Zebrafish Model and the Correlation with Mice.

Polymeric nanocarriers have high biocompatibility for potential drug delivery applications. After entering bloodstream, nanocarriers will circulate, interact with proteins, dissociate, or be cleared by reticuloendothelial system. Zebrafish as a visual animal model, can serve as a tool for screening nanomedicines and monitoring nanocarrier behaviors in vivo. However, a comprehensive correlation between zebrafish and rodent models is currently deficient. Here, different-sized poly(caprolactone) nanocarriers (PCL NCs) are fabricated with or without PEGylation to investigate correlation between zebrafish and mice regarding their biofate via Förster resonance energy transfer technique. Results show that PEGylated PCL NCs have higher integrity in both zebrafish and mice. Small PEG-PCL NCs have longer circulation, while large PEG-PCL NCs have dramatically higher macrophage sequestration in zebrafish and mice spleen, leading to poor circulation. PCL NCs dissociate rapidly with less macrophage sequestration. Moreover, in 7 days postfertilization (dpf) zebrafish, polymers are eliminated via hepatobiliary pathway, which is not fully functional at earlier stages of development. The effects of nanocarrier integrity on macrophage sequestration in zebrafish and good correlation with mice spleen are pioneered to be demonstrated. The findings suggest that 7 dpf zebrafish are suitable as an in vivo screening model of nanocarriers and predict their biofate in rodents.

Cytidine-directed rapid synthesis of water-soluble and highly yellow fluorescent bimetallic AuAg nanoclusters.

Fluorescent gold/silver nanoclusters templated by DNA or oligonucleotides have been widely reported since DNA or oligonucleotides could be designed to position a few metal ions at close proximity prior to their reduction, but nucleoside-templated synthesis is more challenging. In this work, a novel type of strategy taking cytidine (C) as template to rapid synthesis of fluorescent, water-soluble gold and silver nanoclusters (C-AuAg NCs) has been developed. The as-prepared C-AuAg NCs have been characterized by UV-vis absorption spectroscopy, fluorescence, transmission electron microscopy (TEM), energy dispersive X-ray spectroscopy (EDS), X-ray photoelectron spectroscopy (XPS), Fourier transform infrared spectroscopy (FT-IR), and inductively coupled plasma mass spectroscopy (ICP-MS). The characterizations demonstrate that C-AuAg NCs with a diameter of 1.50 ± 0.31 nm, a quantum yield ∼9%, and an average lifetime ∼6.07 µs possess prominent fluorescence properties, good dispersibility, and easy water solubility, indicating the promising application in bioanalysis and biomedical diagnosis. Furthermore, this strategy by rapid producing of highly fluorescent nanoclusters could be explored for the possible recognition of some disease-related changes in blood serum. This raises the possibility of their promising application in bioanalysis and biomedical diagnosis.

The effect of a soft diet on molar dentin formation during the occlusal establishment period.

OBJECTIVE: This study intends to investigate the effect of a soft food diet on molar dentin formation during the occlusal establishment period. It can provide dietary guidance for infants to strengthen their dental structure. DESIGN: 60 BALB/c mice were used to obtain mandibles during lactation (P0.5, P7.5, P15.5, P21.5) and occlusal establishment (P27.5, P33.5, P60.5). The mice were randomly divided into soft or hard diet groups after weaning at day 21.5. Hematoxylin-eosin and aniline blue staining were used to observe the morphology and number of odontoblasts and the amount of molar dentin formation. Immunohistochemistry was performed to observe the proliferation and apoptosis of odontoblasts. The in vivo fluorescence double-labeling was applied to evaluate the rate of molar dentin formation. RESULTS: The soft diet group had poorer periodontal membrane development but more cervical dentin deposition. Alterations in morphology and the number of odontoblasts showed a stronger correlation with age rather than food hardness. There are no significant differences in proliferative and apoptotic behavior of dentin-forming cells between the two groups. Rather, it affected the rate of dentin deposition. The rate of dentin deposition was high in the soft diet group from P21.5 to P27.5, but it was surpassed by the hard diet group within P27.5-P33.5, and the difference between the two groups disappeared at P33.5-P60.5. CONCLUSIONS: A soft diet promotes molar early cervical dentin formation. This advantage is caused by an enhanced odontoblast secretion rate rather than affecting the morphology, number, proliferation, or apoptosis of odontoblasts.

Exploring the release mechanism of micro/nanoplastics from different layers of masks in water: towards reduction of plastic contamination in masks.

During the COVID-19 pandemic, a substantial quantity of disposable face masks was discarded, consisting of three layers of nonwoven fabric. However, their improper disposal led to the release of microplastics (MPs) and nanoplastics (NPs) when they ended up in aquatic environments. To analyze the release kinetics and size characteristics of these masks, release experiments were performed on commercially available disposable masks over a period of 7 days and micro- and nanoplastic releases were detected using fiber counting and nanoparticle tracking analysis. The study's findings revealed that there was no significant difference (p > 0.05) in the quantity of MPs released among the layers of the masks. However, the quantity of NPs released from the middle layer of the mask was 25.9 ± 1.3 × 108 to 81.3 ± 5.3 × 108 particles/piece, significantly higher than the inner and outer layers (p < 0.05). The release process of micro/nanoplastics (M/NPs) from each layer of the mask followed the Elovich equation and the power function equation, indicating that the release was divided into two stages. MPs in the range of 1-500 µm and NPs in the range of 100-300 nm dominated the release from each layer of the mask, accounting for an average of 93.81% and 67.52%, respectively. Based on these findings, recommendations are proposed to reduce the release of M/NPs from masks during subsequent use.

Analysis of the Epidemiologic Characteristics of Children with Hand, Foot and Mouth Disease in China.

Background: We aimed to investigate the epidemiological characteristics of hand, foot and mouth disease (HFMD) and provide recommendations for its control and prevention. Methods: A descriptive epidemiological analysis was designed for HFMD in children through direct network report from 2015 to 2019. Results: From 2015 to 2019, a total 27,395 HFMD cases were reported at Infection Clinic of Shanghai Children's Hospital, Shanghai, China, accounting for 79.24% of the number of reported notifiable infectious disease cases (34,573). The cases number increased since May and reached a peak from June to September, then decreased from October to December with a second small peak in some years. The proportion of cases in boys is higher than in girls (59.7% vs. 40.3%). The cases number of stay-at-home children was higher in 2016 and 2018 (60.07% and 60.95%) than in 2019 (33.47%), and that of kindergarten children and students in 2019 (51.73% and 9.75%) was significantly higher than in other years. Overall, 22606 cases were reported in <5 years group, accounting for 82.52% of the total number of cases during 2015-2019. The proportion of the cases in 5-10 and >10 years groups increased year by year from 2015 to 2019, which is statistically significant. (χ2=71.105, P=0.00; χ2=78.413, P=0.00). Conclusion: The epidemiological characteristics of HFMD had changed during 2015-2019. Analysis of these data can provide helpful evidence to prevention and early treatment of the HFMD, and identification severe cases and handling the outbreaks.

Release of microplastics and nanoplastics in water from disposable surgical masks after disinfection.

During the COVID-19 pandemic, disposable surgical masks were generally disinfected and reused due to mask shortages. Herein, the role of disinfected masks as a source of microplastics (MPs) and nanoplastics (NPs) was investigated. The amount of MPs and NPs released from masks disinfected by UV ranged from 1054 ± 106 to 2472 ± 70 and from 2.55 ± 0.22 × 109 to 6.72 ± 0.27 × 109 particles/piece, respectively, comparable to that of the undisinfected masks, and the MPs were changed to small-sized particles. The amount of MPs and NPs released after alcohol and steam treatment were respectively lower and higher than those from undisinfected masks, and MPs were shifted to small-sized particles. The amount of MPs and NPs released in water after autoclaving was lower than for undisinfected masks. In all, the amount of fibers released after disinfection decreased greatly, and certain disinfection processes were found to increase the amount of small-sized NPs released from masks into aqueous environments.

Release kinetics of microplastics from disposable face masks into the aqueous environment.

Disposable face masks are widely used as primary personal protective equipment to control the spread of the SARS-CoV-2 virus. Disposable face masks have been identified as a source of microplastics and a new threat to the environment when improperly handled. To understand the release of microplastics from discarded masks into water, the release quantities of microplastics from three types of disposable face masks (N95, medical surgical, and normal medical masks) were measured within 24 h and their release kinetics were analyzed over seven days. Results showed that polypropylene microplastics fibers and debris of various colors were released. N95 masks released 801 ± 71-2667 ± 97 microplastic particles/(piece·d), medical surgical masks released 1136 ± 87-2343 ± 168 microplastic particles/(piece·d), and normal medical masks released 1034 ± 119-2547 ± 185 microplastic particles/(piece·d), irrespective of the price, weight, or type of mask. The microplastics were first released fast and then slow. The Elovich equation described the release kinetics (R2 > 0.990), and the release rate did not differ with the type of mask. Microplastics of 100-500 µm and of <100 µm were released in large quantities and at rapid rates. Fiber and transparent microplastics accounted for a large proportion of those released, and their daily release proportion increased with time. Fiber microplastics <500 µm in length were predominant in the microplastics released from disposable face masks, indicating that disposable face masks could be a critical source of these in the aqueous environment. There is an urgent need to take action to implement a waste management system limiting the number of masks entering the environment.

Apoptosis-Sensing Xenograft Zebrafish Tumor Model for Anticancer Evaluation of Redox-Responsive Cross-Linked Pluronic Micelles.

A suitable animal model for preclinical screening and evaluation in vivo could vastly increase the efficiency and success rate of nanomedicine development. Compared with rodents, the transparency of the zebrafish model offers unique advantages of real-time and high-resolution imaging of the whole body and cellular levels in vivo. In this research, we established an apoptosis-sensing xenograft zebrafish tumor model to evaluate the anti-cancer effects of redox-responsive cross-linked Pluronic polymeric micelles (CPPMs) visually and accurately. First, doxorubicin (Dox)-loaded CPPMs were fabricated and characterized with glutathione (GSH)-responsive drug release. Then, the B16F10 xenograft zebrafish tumor model was established to mimic the tumor microenvironment with angiogenesis and high GSH generation for redox-responsive tumor-targeting evaluation in vivo. The high GSH generation was first verified in the xenograft zebrafish tumor model. Compared with ordinary Pluronic polymeric micelles, Dox CPPMs had a much higher accumulation in zebrafish tumor sites. Finally, the apoptosis-sensing B16F10-C3 xenograft zebrafish tumor model was established for visual, rapid, effective, and noninvasive assessment of anti-cancer effects at the cellular level in vivo. The Dox CPPMs significantly inhibited the proliferation of cancer cells and induced apoptosis in the B16F10-C3 xenograft zebrafish tumor model. Therefore, the redox-responsive cross-linked Pluronic micelles showed effective anti-cancer therapy in the xenograft zebrafish tumor model. This xenograft zebrafish tumor model is available for rapid screening and assessment of anti-cancer effects in preclinical studies.

Effects of polymer aging on sorption of 2,2',4,4'-tetrabromodiphenyl ether by polystyrene microplastics.

The sorption behavior of 2,2',4,4'-tetrabromodiphenyl ether (BDE-47) on aged polystyrene (PS) microplastics via seawater soaking, ultraviolet (UV) irradiation, seawater soaking and UV irradiation together was investigated. The effects of environmental factors including salinity, pH, and dissolved organic matter (DOM) on sorption of BDE-47 by the aged PS microplastics were analyzed. The equilibrium sorption capacity of BDE-47 by virgin PS, aged PS with seawater soaking, aged PS with UV irradiation and aged PS with seawater soaking and UV irradiation was 6.16, 4.96, 3.53, and 3.75 ng/g, respectively. The decrease in sorption capacity was related to the increase of surface crystallinity and the appearance of surface oxygen-containing functional groups. The kinetic and isotherm models suggested that aging did not change the sorption mechanism of BDE-47 on PS microplastics. pH had negligible impacts on BDE-47 sorption by virgin and aged PS. Sorption capacity of BDE-47 on aged PS in water with high salinity was lower than that on virgin PS, and DOM has less effect on the sorption of BDE-47 on aged PS.

Toward understanding the prolonged circulation and elimination mechanism of crosslinked polymeric micelles in zebrafish model.

Understanding the behaviors of nanomedicines in vivo is one of the most important prerequisites for the design and optimization of nanomedicines. However, the in vivo tracking of nanomedicines in rodents is severely limited by the restricted imaging possibilities within these animals. To meet these needs, the FRET (fluorescence or Förster resonance energy transfer) imaging combined with visual zebrafish larvae model (7 dpf) was used to study the behaviors of polymeric micelles in vivo at high spatiotemporal resolution. Firstly, the FRET ordinary Pluronic micelles (OPMs) and disulfide bond crosslinked Pluronic micelles (CPMs) were synthesized to quantify their integrity in vitro and in vivo by FRET ratio. The behaviors and integrity of OPMs and CPMs in vivo were visually investigated in zebrafish larvae across the entire living organism and at cellular molecular level after intravenous microinjection. Results showed that OPMs were rapidly disassociated in circulation, then largely sequestrated by the endothelial cells (ECs) of caudal vein (CV) and liver in zebrafish larvae, which resulted in quick elimination from blood circulation. While the CPMs were more stable and escaped the sequestration by ECs of CV and liver, which prolonged their circulation in blood. Moreover, we pioneered to use the zebrafish model to reveal that polymeric micelles were eliminated through hepatobiliary pathway after disassociation. While the intact micelles were relatively difficult to eliminate. We further verified that the scavenger receptors of ECs but not the macrophages mainly mediated the elimination of polymeric micelles in CV and liver of zebrafish larvae. These finding on behaviors and elimination mechanisms of polymeric micelles in zebrafish model could contribute to the rational design and optimization of nanomedicines, further guide their studies in rodents.

Tensile force produced by a lipoma in the masseteric space possibly causing hyperostosis of the angle of the mandible.

BACKGROUND: A lipoma is a benign tumor which may occur in the adipose tissue of any part of the body. The tumor is most commonly found on the trunk and extremities. Although it is the most common tumor of mesenchymal origin in the head and neck, its incidence is relatively rare. Lipoma of the head and neck is usually located in subcutaneous tissue. It is rarely deep seated with osseous involvement and rarely occurs in children, especially below the age of 10. CASE REPORT: The case of a painless mass of one-year history in the right parotidomasseteric region of an eight-year-old boy is presented. The mass was revealed to be a classical lipoma in the masseteric space, possibly causing hyperostosis of the angle of the mandible. After removal of the mass and a spherical protuberance in the angle of the mandible, the boy recovered and no recurrence was noted after one-year follow-up. CONCLUSIONS: Hyperostosis is a rare phenomenon with lipoma. In this case, the hypothesis was posed that the tensile force produced by the lipoma in the masseteric space possibly caused hyperostosis of the angle of the mandible. To the authors' knowledge, a classical lipoma arising from the deep fascial space resulting in osseous change has not been described in the literature before.

Sorption of polybrominated diphenyl ethers by microplastics.

The sorption of polybrominated diphenyl ethers (PBDEs) onto polyethylene (PE), polypropylene (PP), polystyrene (PS), and polyamide (PA) microplastics was analyzed using different kinetic and isotherm models, and under various environmental conditions, including temperature, pH, salinity and dissolved organic matter (DOM). The sorption capacity was in the order of PS > PA > PP > PE, due to the different crystallinity, specific surface area, and surface structure. PS demonstrated the equilibrium sorption capacity, namely, 6.41 ng/g BDE-47, 12.83 ng/g BDE-99, and 14.42 ng/g BDE-153. The second-order kinetic model described the sorption kinetics of PBDEs, and surface sorption was the main mechanism. The sorption of PBDEs by microplastics was a multilayer and physical process. Low temperatures reduced BDE-47 sorption on microplastics, and sorption was a spontaneous and endothermic process. The sorption of BDE-47 was not significantly influenced by pH and salinity. However, DOM exerted a negative effect on the sorption of BDE-47.

Immobilized lignin peroxidase on Fe3O4@SiO2@polydopamine nanoparticles for degradation of organic pollutants.

Lignin peroxidase (LiP) was obtained from Pichia methanolica through heterologous expression. LiP was extracted, purified, and immobilized on Fe3O4@SiO2@polydopamine (PDA) nanoparticles to acquire immobilized LiP. The optimal preparation conditions for immobilized LiP were investigated. Results showed that the immobilization efficiency of immobilized LiP reached 56.37% when the enzyme amount, PDA concentration, and immobilization time were 12 mg, 1.6 mg/mL, and 12 h, respectively. Compared with free LiP, the immobilized LiP showed good thermal stability and storage stability and improved pH tolerance. It also retained more than 30% of its initial activity after 8 cycles, demonstrating its improved reusability. The immobilized LiP demonstrated efficacy of reaction of 100%, 100%, 100%, 100%, 79%, 73%, and 65% for tetracycline, dibutyl phthalate, 5-chlorophenol, phenol, phenanthrene, fluoranthene, and benzo(a)pyrene, respectively, while the inactivated immobilized LiP only adsorbed <25% of phenanthrene and fluoranthene. The dissipation of organic pollutants was a combination of degradation and adsorption, with the former playing a more important role.

Molecular cloning and transcriptional expression analysis of an intracellular beta-glucosidase, a family 3 glycosyl hydrolase, from the edible straw mushroom, Volvariella volvacea.

A beta-glucosidase, with a molecular mass of 95 kDa, was isolated from extracts of Volvariella volvacea mycelium grown on crystalline cellulose. Degenerate primers based on the N-terminal sequences of purified beta-glucosidase and two protease-generated peptides were used to generate cDNA fragments encoding a portion of the beta-glucosidase gene (bgl), and rapid amplification of cDNA ends was used to obtain full-length cDNA clones. The cDNA of bgl contained an ORF of 2586 bp coding for 862 amino acids. Alignment of the deduced amino-acid sequence of beta-glucosidase with deduced amino acid sequences of other microbial beta-glucosidases showed the highest overall homology with glycoside hydrolase family 3 beta-glucosidases from fungi. Transcripts of bgl were detected in total RNA extracted from mycelium grown on cellulose and cellobiose, and from mycelium pre-grown for 72 h in basal medium containing 1% (w/v) sorbitol following addition of alpha-lactose, beta-lactose, cellobiose, d- xylose, l-sorbose, beta-gentiobiose, sophorose or d-galactose. Addition of l-sorbose and d-glucosamine to mycelium grown on 1% (w/v) crystalline cellulose greatly increased the level of bgl expression. bgl Was expressed at various stages of the mushroom developmental cycle (substrate colonization to mature fruit body), although the number of bgl transcripts in pinhead and button stages was slightly smaller.

Complete in vitro oogenesis: retrospects and prospects.

Precise control of mammalian oogenesis has been a traditional focus of reproductive and developmental biology research. Recently, new reports have introduced the possibility of obtaining functional gametes derived in vitro from stem cells. The potential to produce functional gametes from stem cells has exciting applications for regenerative medicine though still remains challenging. In mammalian females ovulation and fertilization is a privilege reserved for a small number of oocytes. In reality the vast majority of oocytes formed from primordial germ cells (PGCs) will undergo apoptosis, or other forms of cell death. Removal occurs during germ cell cyst breakdown and the establishment of the primordial follicle (PF) pool, during the long dormancy at the PF stage, or through follicular atresia prior to reaching the ovulatory stage. A way to solve this limitation could be to produce large numbers of oocytes, in vitro, from stem cells. However, to recapitulate mammalian oogenesis and produce fertilizable oocytes in vitro is a complex process involving several different cell types, precise follicular cell-oocyte reciprocal interactions, a variety of nutrients and combinations of cytokines, and precise growth factors and hormones depending on the developmental stage. In 2016, two papers published by Morohaku et al. and Hikabe et al. reported in vitro procedures that appear to reproduce efficiently these conditions allowing for the production, completely in a dish, of a relatively large number of oocytes that are fertilizable and capable of giving rise to viable offspring in the mouse. The present article offers a critical overview of these results as well as other previous work performed mainly in mouse attempting to reproduce oogenesis completely in vitro and considers some perspectives for the potential to adapt the methods to produce functional human oocytes.

[Expression of c-fos, OPG, OPGL in rabbit mandibular distraction osteogenesis zone].

OBJECTIVE: To evaluate the possible signal transduction mechanism of the mechanical stress induced by the distraction procedure in osteocytes. METHODS: An animal model of mandibular distraction osteogenesis in rabbits was established. The expressions of c-fos, OPG and OPGL were detected by ultrasensitive S-P immunohistochemical method. RESULT: At 4 and 8 days after distraction, distraction zone showed strong positive staining of c-fos, which were apparently higher than that in distraction zone of 2, 4 and 6 weeks after consolidation. At 4 and 8 days after distraction and 2 weeks after consolidation, the expression of OPG was strong, and then wore off gradually at 4 and 6 weeks after consolidation. Weak signals of OPGL could be detected at 6 weeks after consolidation only. CONCLUSION: c-fos, OPG and OPGL are important regulators in distraction osteogenesis. c-fos is interrelated with the mechanical stress induced by the distraction procedure closely, OPG promotes new bone formation, while OPGL plays a more active role in bone remodeling.

Polybrominated diphenyl ethers in dissolved and suspended phases of seawater and in surface sediment from Jiaozhou Bay, North China.

The concentration and distribution of polybrominated diphenyl ethers (PBDEs) in the dissolved phase (DP) and suspended particulate matter (SPM) of seawater and in surface sediment obtained from Jiaozhou Bay (JZB) were determined. The potential sources of these compounds were evaluated. The total concentrations of 14 PBDEs (∑14PBDE) in DP and SPM were 0.09-1.35 and 1.17-5.45ng/L, respectively, indicating that the PBDE congeners are predominantly partitioned into the SPM fraction. The ∑14PBDE concentrations in sediment ranged from 2.18ng/g to 10.59ng/g with a mean value of 6.59ng/g. BDE-209 was dominant among 14 PBDE congeners, and BDE-47 was another abundant congener in the SPM and sediment samples. The PBDE concentration in SPM showed a significantly positive correlation with chlorophyll a level (r(2)=0.496, p<0.05), and that in the sediment exhibited a significantly positive correlation with clay proportion (r(2)=0.846, p<0.01). This result suggests that the phytoplankton or debris in the suspended particulates play an important role in PBDE accumulation and transportation and that PBDEs are preferentially enriched in clays in the sediment. PBDE concentrations were higher in the inner and mouth regions than in the outer and middle regions, and decreased with water depth in the water column. Non-parametric multidimensional scaling ordination showed that BDE-209 and BDE-47 are the important products discriminating PBDE contamination. This result is associated with industrial waste discharge from urban areas and with heavy ship traffic, indicating that DecaBDE and PentaBDE products are the potential sources of PBDEs. PBDE concentrations were lower in the sediment in JZB than in other coastal areas in South China, although the PBDE concentrations in DP were higher than those in some coastal areas worldwide. Moreover, the PBDE concentrations in DP and sediment increased in the last decade.

[Transforming growth factor-beta1-loaded fibrin sealant promote bone marrow Mesenchymal stem cells to contract injectable tissue engineering cartilage in vivo].

OBJECTIVE: To investigate the feasibility that transforming growth factor-beta1 (TGF-beta1) -loaded fibrin sealant (FS) promotes bone marrow mesenchymal stem cells (BMSCs) to create tissue engineering cartilage in vivo. METHODS: The BMSCs were isolated from healthy human and amplified in vitro, and then induced by defined medium containing TGF-beta1 and dexamethasone. After 7 days the induced BMSCs were collected and mixed with TGF-beta1-loaded FS or FS as BMSCs+ FS-TGF-beta1 group and BMSCs+ FS experimental group. Then the mixture was injected by a needle into the dorsum of nude mice. In control group, only FS or BMSCs were injected. The tissue engineering specimens were harvested from nude mice 12 weeks later. Gross observation, average wet weight measurement, glycosaminoglycan (GAG) quantification, histology and immunohistochemistry were used to evaluate the results. RESULTS: The BMSCs have possessed the shape and functional characters of chondrocyte when transferred to a defined medium. After injection of the mixture, the cartilage-like tissue were formed in two experimental groups. Compared with BMSC+ FS group, the specimens of BMSCs +FS-TGF-beta1 group were larger and firmer. Alcian staining showed better metachromatic matrix formation. The GAG contents were significantly higher. Immunohistochemical staining of collagen type II was stronger. However, no cartilage-like tissue was formed in two control groups. CONCLUSION: TGF-beta1-loaded FS can promote BMSCs to contract injectable tissue engineering cartilage in vivo.

[Preparation and antitumor immunity of long circulating nano-liposome encapsulated tumor specific antigen].

AIM: To prepare Nano-Liposome encapsulated MAGE3/HSP70(NL M3H) and study its character and antitumor immunity in mouse. METHODS: NL M3H was prepared by the thin film-dispersion ultrasonic. The shape and size of NL M3H were detected by electron microscope. The encapsulation rate, drug-carrying capacity, stability and the releasing character were tested by Sephedex-G100 gel filtration. The mouse was immunized by NL M3H, and the antitumor immunity was detected by ELISPOT and LDH release assay. RESULTS: The mean size of NL M3H was lower than 100 nm. The encapsulation rate was 38%.The drug content was 0.038 g/L. NL M3H has good stability after stored in 4 degrees C for 6 months. The releasing profile showed that 74 percent of proteins was released during the first 24 hours in saline. The results of ELISPOT and LDH release assay showed that NL M3H generated tumor specific cytotoxic T lymphocyte(CTL)to damage tumor cell. CONCLUSION: NL M3H has novel characters, it can generate specific CTL to kill tumor cell, and can be used as new kind of vaccine against tumor.