Transdermal microarrayed electroporation for enhanced cancer immunotherapy based on DNA vaccination.

Despite the tremendous clinical potential of nucleic acid-based vaccines, their efficacy to induce therapeutic immune response has been limited by the lack of efficient local gene delivery techniques in the human body. In this study, we develop a hydrogel-based organic electronic device (µEPO) for both transdermal delivery of nucleic acids and in vivo microarrayed cell electroporation, which is specifically oriented toward one-step transfection of DNAs in subcutaneous antigen-presenting cells (APCs) for cancer immunotherapy. The µEPO device contains an array of microneedle-shaped electrodes with pre-encapsulated dry DNAs. Upon a pressurized contact with skin tissue, the electrodes are rehydrated, electrically triggered to release DNAs, and then electroporate nearby cells, which can achieve in vivo transfection of more than 50% of the cells in the epidermal and upper dermal layer. As a proof-of-concept, the µEPO technique is employed to facilitate transdermal delivery of neoantigen genes to activate antigen-specific immune response for enhanced cancer immunotherapy based on a DNA vaccination strategy. In an ovalbumin (OVA) cancer vaccine model, we show that high-efficiency transdermal transfection of APCs with OVA-DNAs induces robust cellular and humoral immune responses, including antigen presentation and generation of IFN-γ+ cytotoxic T lymphocytes with a more than 10-fold dose sparing over existing intramuscular injection (IM) approach, and effectively inhibits tumor growth in rodent animals.

The tendency of anemia of inflammation in periodontal diseases.

Anemia of inflammation (AI) is associated with inflammatory diseases, and inflammation-induced iron metabolism disorder is the major pathogenic factor. Earlier studies have reported a tendency of AI in periodontitis patients, but the explicit relationship and possible pathological mechanisms remain unclear. Here, the analyses of both periodontitis patients and a mouse model of ligature-induced experimental periodontitis showed that periodontitis was associated with lower levels of hemoglobin and hematocrit with evidence of systemic inflammation (increased white blood cell levels) and evidence of iron restriction (low serum iron along with a high serum hepcidin and ferritin levels), in accordance with the current diagnosis criteria for AI. Moreover, periodontal therapy improved the anemia status and iron metabolism disorders. Furthermore, the increased level of hepcidin and significant correlation between hepcidin and key indicators of iron metabolism emphasized the pivotal role of hepcidin in the pathogenesis of periodontitis-related AI. Administration of the signal transducer and activator of transcription 3 (STAT3) inhibitors Stattic suggested that the IL-6-STAT3-hepcidin signaling pathway participated in this regulatory process. Together, these findings demonstrated that periodontitis should be considered an inflammatory disease that contributes to the development of AI; furthermore, IL-6-STAT3-hepcidin signaling pathway plays a key regulatory role in the pathogenesis of periodontitis-related AI. Our study will provide new insights into the systemic effects of periodontitis, while meaningfully expanding the spectrum of inflammatory diseases that contribute to AI.

CdSe/TiO2NTs Heterojunction-Based Nonenzymatic Photoelectrochemical Sensor for Glucose Detection.

Compared with a single semiconductor, the heterojunction formed by two different semiconductors usually has higher light utilization and better photoelectric performance. By using stable TiO2 nanotubes as the main subject, CdSe/TiO2NTs heterojunctions were synthesized by a hydrothermal method. XRD, TEM, SEM, PL, UV-vis, and EIS were used to characterize the fabricated CdSe/TiO2NTs. Under visible light irradiation, CdSe/TiO2NTs heterojunctions exhibited a higher absorption intensity and lower degree of photogenerated carrier recombination than TiO2. The electrons and holes were proven to be effectively separated in this heterojunction via theoretical calculation. Under CdSe/TiO2NTs' optimal conditions, the glucose concentrations (10-90 µM) had a linear relationship with the photocurrent value, and the detection limit was 3.1 µM. Moreover, the CdSe/TiO2NTs sensor exhibited good selectivity and stability. Based on the experimental data and theoretical calculations, its PEC sensing mechanism was also illuminated.

Multicopy Chromosome Integration and Deletion of Negative Global Regulators Significantly Increased the Heterologous Production of Aborycin in Streptomyces coelicolor.

Aborycin is a type I lasso peptide with a stable interlocked structure, offering a favorable framework for drug development. The aborycin biosynthetic gene cluster gul from marine sponge-associated Streptomyces sp. HNS054 was cloned and integrated into the chromosome of S. coelicolor hosts with different copies. The three-copy gul-integration strain S. coelicolor M1346::3gul showed superior production compared to the one-copy or two-copy gul-integration strains, and the total titer reached approximately 10.4 mg/L, i.e., 2.1 times that of the native strain. Then, five regulatory genes, phoU (SCO4228), wblA (SCO3579), SCO1712, orrA (SCO3008) and gntR (SCO1678), which reportedly have negative effects on secondary metabolism, were further knocked out from the M1346::3gul genome by CRISPR/Cas9 technology. While the ΔSCO1712 mutant showed a significant decrease (4.6 mg/L) and the ΔphoU mutant showed no significant improvement (12.1 mg/L) in aborycin production, the ΔwblA, ΔorrA and ΔgntR mutations significantly improved the aborycin titers to approximately 23.6 mg/L, 56.3 mg/L and 48.2 mg/L, respectively, which were among the highest heterologous yields for lasso peptides in both Escherichia coli systems and Streptomyces systems. Thus, this study provides important clues for future studies on enhancing antibiotic production in Streptomyces systems.

Highly Sensitive Temperature-Pressure Bimodal Aerogel with Stimulus Discriminability for Human Physiological Monitoring.

Multimodal sensor with high sensitivity, accurate sensing resolution, and stimuli discriminability is very desirable for human physiological state monitoring. A dual-sensing aerogel is fabricated with independent pyro-piezoresistive behavior by leveraging MXene and semicrystalline polymer to assemble shrinkable nanochannel structures inside multilevel cellular walls of aerogel for discriminable temperature and pressure sensing. The shrinkable nanochannels, controlled by the melt flow-triggered volume change of semicrystalline polymer, act as thermoresponsive conductive channels to endow the pyroresistive aerogel with negative temperature coefficient of resistance of -10.0% °C-1 and high accuracy within 0.2 °C in human physiological temperature range of 30-40 °C. The flexible cellular walls, working as pressure-responsive conductive channels, enable the piezoresistive aerogel to exhibit a pressure sensitivity up to 777 kPa-1 with a detectable pressure limit of 0.05 Pa. The pyro-piezoresistive aerogel can detect the temperature-dependent characteristics of pulse pressure waveforms from artery vessels under different human body temperature states.

Legal determination of surrogate child parenthood in China.

Commercial surrogacy is neither prohibited nor approved in China. As the underground surrogacy market keeps expanding fast, the disputes over parenthood of surrogate child occur more and more frequently. Since there are no specific rules for surrogacy, Chinese courts have to make legal determination by applying current laws. This article firstly analyses the contradictions of existing laws and regulations on natural parenthood when apply them in the cases of surrogacy. In China, since the natural parenthood is built on reproductive connections between parents and child, this article provides a typology of surrogacy in terms of the reproductive role each party has played. By combining it with the case study, the judicial views on different issues are presented. This article further discusses the inadequacies of current legal determination of parenthood, ethical controversies in surrogacy and recommends some suggestions on regulating the surrogacy and protecting the rights and interests of women and children.

Investigation of electron transfer between single plasmon and graphene by dark field spectroscopy.

We investigated the electron transfer time between single plasmonic gold nanoparticles and graphene with our home-build spectral imaging dark-field microscope. The process of electron transfer is supposed to be shuttling of hot electrons on the nanoparticle-graphene interface, resulting in a slight broadening of the scattering spectrum. For detecting the minor spectrum broadening, we firstly characterized our setup systematically and then calibrated its intrinsic error. We found the mechanism of a common but normally neglected setup error, scattering spectrum broadening, which is caused by the bandwidth of the incident light and could exist in most fast dark-field microscopy setups. We corrected the linewidth of plasmon scattering spectra theoretically by both numerical and analytical solution, and then realized it experimentally by tuning the bandwidth of the incident light. After calibration, we revisited scattering spectra of 700 small aspect ratio nanorods on glass and monolayer graphene revealing a typical 14.3 meV linewidth broadening. Furthermore, we measured four other kinds of gold nanoparticles on glass, mono- and bilayer graphene for deeper understanding of the electron transfer. A common linewidth broadening is found for each kind of particle agreeing well with previous theory. However, an unconventional linewidth narrowing is also discovered for big particles whose resonance wavelength is close to the near infrared region. It implies a competitive mechanism in the electron transfer process which could not only increase the damping of small particles, causing a linewidth broadening, but also simplify the electric field pattern for big particles, leading to a linewidth narrowing, according to our Mie theory simulation.

Digital Workflow for Full-Arch Immediate Implant Placement Using a Stackable Surgical Guide Fabricated Using SLM Technology.

The failing dentition of partially edentulous individuals may be used as an initial reference for stackable restrictive surgical guides during full-arch immediate implant placement. The stackable guide option derived from a digital workflow increases the predictability of the performance of bone reduction, immediate implant placement, and immediate loading of provisional implant-supported fixed dental prostheses. The present paper aims to report a practical approach to design and produce a metal framework with occlusal rests to facilitate the use of a tooth-supported surgical guide when full-arch immediate implant placement is indicated in patients with failing dentition.

3D Hydrogen Titanate Nanotubes on Ti Foil: A Carrier for Enzymatic Glucose Biosensor.

Glucose oxidase (GOx) based biosensors are commercialized and marketed for the high selectivity of GOx. Incorporation nanomaterials with GOx can increase the sensitivity performance. In this work, an enzyme glucose biosensor based on nanotubes was fabricated. By using Ti foil as a carrier, hydrogen titanate nanotubes (HTNTs), which present fine 3D structure with vast pores, were fabricated in-situ by the hydrothermal treatment. The multilayer nanotubes are open-ended with a diameter of 10 nm. Then glucose oxidase (GOx) was loaded on the nanotubes by cross-linking to form an electrode of the amperometric glucose biosensor (GOx/HTNTs/Ti electrode). The fabricated GOx/HTNTs/Ti electrode had a linear response to 1-10 mM glucose, and the response time was 1.5 s. The sensitivity of the biosensor was 1.541 µA·mM-1·cm-2, and the detection limit (S/N = 3) was 59 µM. Obtained results indicate that the in-situ fabrication and unique 3D structure of GOx/HTNTs/Ti electrode are beneficial for its sensitivity.

Sensitive fluorescence assay of organophosphorus pesticides based on the fluorescence resonance energy transfer between CdTe quantum dots and porphyrin.

A sensitive and selective quantum dot (QD)-based fluorescence resonance energy transfer (FRET) biosensor was successfully fabricated for the detection of organophosphorus pesticides (OPs). 5,10,15,20-Tetra(4-pyridyl)porphyrin (TPyP) with meso-pyridyl substituents was bound to the surface of CdTe QDs to produce self-assembled nanosensors, and the process of FRET between QDs and TPyP occurred. However, the process of FRET was switched off with the addition of OPs, due to the combination between TPyP and OPs. The fluorescence intensity of TPyP (donor) would decrease gradually with the increasing concentration of OPs. Under optimal conditions, a linear correlation was established between the fluorescence intensity ratio ITPyP/IQDs and the concentration of paraoxon in the range of 9.09 × 10(-12)-1.09 × 10(-6) mol L(-1) with a detection limit of 3.15 × 10(-12) mol L(-1). The attractive sensitivity was obtained due to the efficient FRET and the superior fluorescence properties of QDs. The proposed method was successfully applied to the determination of the OPs in real fruit samples with satisfactory results.

Highly-sensitive organophosphorus pesticide biosensors based on CdTe quantum dots and bi-enzyme immobilized eggshell membranes.

An optical biosensing method using CdTe quantum dots (QDs) and bi-enzyme-immobilized eggshell membranes for the determination of organophosphorus pesticides (OPs) has been developed. Increasing amounts of OPs led to a decrease of the enzymatic activity and thus a decrease in the production of hydrogen peroxide (H2O2), which can quench the fluorescence of the CdTe QDs. Under the optimum conditions, there was a good linear relationship between the enzyme inhibition percentage and the logarithm of paraoxon or parathion concentration in the range of 1.0 × 10(-11)-1.0 × 10(-6) mol L(-1). The detection limit (S/N = 3) of the proposed biosensors were as low as 4.30 × 10(-12) mol L(-1) for paraoxon and 2.47 × 10(-12) mol L(-1) for parathion. The bi-enzyme-immobilized eggshell membrane demonstrated a long shelf-life of at least 2 months and the results showed good repeatability. The proposed method was successfully applied to the determination of the OPs in real fruit samples with satisfactory results.

Expression and identification of the ADF-linker-3-1E gene of Eimeria acervulina of chicken.

Coccidiosis is a widely distributed disease with higher mortality and morbidity, which is caused by several species of protozoan parasites belonging to the genus Eimeria and recognized as a serious challenge for the poultry industry. This research was conducted to construct the recombinant plasmid pET32a(+)-ADF-linker-3-1E of Eimeria acervulina (E. acervulina) of the chicken and test the bioactivity of the ADF-linker-3-1E protein. The ADF-linker-3-1E gene of E. acervulina of the chicken was cloned by splicing by overlap extension by the polymerase chain reaction (SOE-PCR) and then inserted into the pET32a(+) to construct the recombinant plasmid pET32a(+)-ADF-linker-3-1E. The recombinant plasmid was transformed into Escherichia coli Rosetta (DE3) competent cells and then induced by IPTG (0.6 mmol/L). The expressed product in the culture medium was identified by the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The bioactivity of the ADF-linker-3-1E protein was tested by Western blotting. The result showed that the amplified ADF-linker-3-1E gene was about 1346 bp. The PCR amplification with the recombinant plasmid pET-32a(+)-ADF-linker-3-1E as a template resulted in a special band of 1346 bp. The digested products resulted in two fragments of 1346 bp target fragment and 5.9 kb pET-32a(+)-vector fragment. The results indicated that the ADF-linker3-1E gene was successfully inserted into the pET-32a(+)-vector. The expressed products in the culture medium resulted in a single band of approximately 54.8 kDa by SDS-PAGE. Western blotting analysis indicated that the recombinant protein could be reacted specifically with His-Tag(2A8) Mouse mAb. This study indicated that the ADF-linker-3-1E protein with good bioactivity was successfully obtained, which laid a foundation for the exploitation of the nuclear vaccine by using the ADF-linker-3-1E protein.

Synchrotron X-ray tomographic microscopy of fossil embryos.

Fossilized embryos from the late Neoproterozoic and earliest Phanerozoic have caused much excitement because they preserve the earliest stages of embryology of animals that represent the initial diversification of metazoans. However, the potential of this material has not been fully realized because of reliance on traditional, non-destructive methods that allow analysis of exposed surfaces only, and destructive methods that preserve only a single two-dimensional view of the interior of the specimen. Here, we have applied synchrotron-radiation X-ray tomographic microscopy (SRXTM), obtaining complete three-dimensional recordings at submicrometre resolution. The embryos are preserved by early diagenetic impregnation and encrustation with calcium phosphate, and differences in X-ray attenuation provide information about the distribution of these two diagenetic phases. Three-dimensional visualization of blastomere arrangement and diagenetic cement in cleavage embryos resolves outstanding questions about their nature, including the identity of the columnar blastomeres. The anterior and posterior anatomy of embryos of the bilaterian worm-like Markuelia confirms its position as a scalidophoran, providing new insights into body-plan assembly among constituent phyla. The structure of the developing germ band in another bilaterian, Pseudooides, indicates a unique mode of germ-band development. SRXTM provides a method of non-invasive analysis that rivals the resolution achieved even by destructive methods, probing the very limits of fossilization and providing insight into embryology during the emergence of metazoan phyla.

Environmental significance of atmospheric nitrogen deposition in the transition zone between the Tibetan Plateau and arid region.

The ecological effect of atmospheric N deposition has become a hot research point along with intensive human activities and global climatic change. As the transition zone between the Tibetan Plateau and the arid region, the Qilian Mountains are important ecological barriers and source regions of inland rivers in northwest China. However, the quantification of N deposition in the transition zone between the Tibetan Plateau (TP) and the arid region remains unclear, primarily due to the lack of in situ measurements. Hence, an observational study was conducted on the Qilian Mountains, and precipitation data were collected. Approximately 1382 samples were collected and analyzed regarding their characteristics and environmental effects of the atmospheric N wet deposition. The annual wet deposition of atmospheric N was 10.05 kg/hm2, and showed a decreasing trend from the south to the north of the Qilian Mountains. NO3--N deposition was the main form of wet deposition of atmospheric N on the Qilian Mountains, accounting for 73.83% of the DIN deposition. Additionally, altitude, meteorological factors, and ecosystem types were found to influence the wet deposition of atmospheric N. The contribution of NO3--N to the wet deposition of atmospheric N in meadows, forests, grasslands, farmlands, and towns was 48.38%, 71.55%, 77.54%, 69.61%, and 82.84%, respectively. This study provides a scientific basis for the effective management and sustainable development of environmental protection in the transition zone between the TP and the arid region. The results showed that relevant policies, as well as scientific and governmental measures, could contribute to reducing N deposition effectively. However, the further mitigation measures should be proposed and strictly enforced.

Bronchial Artery Chemoembolization for Hemoptysis in Advanced Primary Lung Cancer.

BACKGROUND: To evaluate the safety and effectiveness of bronchial arterial infusion chemoembolization (BAICE) for lung cancer with hemoptysis. PATIENTS AND METHODS: Retrospectively analyze clinical data of patients undergoing BAICE for the treatment of lung cancer with hemoptysis, evaluate the clinical efficacy of this approach, observe postoperative adverse reactions, and analyze hemoptysis-free survival (HFS) and overall survival (OS). RESULTS: All 187 patients underwent BAICE with technical success rate of 100%, clinical success rate of 86.6%, clinical failure rate of 13.4%. After BAICE, the tumor curative effect was evaluated as complete remission in 56 cases, partial remission in 82 cases, stable disease in 26 cases, and progressive disease in 6 cases. The objective response rate was 73.8%, and the disease control rate was 87.7%. Median HFS of the 154 clinically successful cases was 10.5 months (95% confidence interval [CI]: 8.11-11.89). The degree of hemoptysis (massive hemoptysis hazard ratio [HR] = 5.9, 95% CI: 3.43-10.16, P = .00) and cavitary lung mass (HR = 2.39, 95% CI: 1.44-3.99, P = .001), were significantly related to a reduction in HFS after BAICE. The 6-month and 12-month survival rates were 66.5% and 45.6%, respectively. The median OS of clinically successful cases was 13.0 months (95% CI: 11.22-14.77). The median OS of 16 clinically failed cases was 2.0 months (95% CI: 0.41-2.45) (P < .001). All adverse events were grade 1. CONCLUSION: BAICE for advanced lung cancer with hemoptysis is safe, effective, and tolerable.

Tailoring of the photocatalytic activity of CeO2 nanoparticles by the presence of plasmonic Ag nanoparticles.

The present study investigates basic features of a photoelectrochemical system based on CeO2 nanoparticles fixed on gold electrodes. Since photocurrent generation is limited to the absorption range of the CeO2 in the UV range, the combination with metal nanoparticles has been studied. It can be shown that the combination of silver nanoparticles with the CeO2 can shift the excitation range into the visible light wavelength range. Here a close contact between both components has been found to be essential and thus, hybrid CeO2@Ag nanoparticles have been prepared and analyzed. We have collected arguments that electron transfer occurs between both compositional elements of the hybrid nanoparticles.The photocurrent generation can be rationalized on the basis of an energy diagram underlying the necessity of surface plasmon excitation in the metal nanoparticles, which is also supported by wavelength-dependent photocurrent measurements. However, electrochemical reactions seem to occur at the CeO2 surface and consequently, the catalytic properties of this material can be exploited as exemplified with the photoelectrochemical reduction of hydrogen peroxide. It can be further demonstrated that the layer-by layer technique can be exploited to create a multilayer system on top of a gold electrode which allows the adjustment of the sensitivity of the photoelectrochemical system. Thus, with a 5-layer electrode with hybrid CeO2@Ag nanoparticles submicromolar hydrogen peroxide concentrations can be detected.

CdS-Modified TiO2 Nanotubes with Heterojunction Structure: A Photoelectrochemical Sensor for Glutathione.

The formation of heterojunction structures can effectively prevent the recombination of photogenerated electron-hole pairs in semiconductors and result in the enhancement of photoelectric properties. Using TiO2 nanotubes (prepared using the hydrothermal-impregnation method) as carriers, CdS-TiO2NTs were fabricated as a photoelectrochemical (PEC) sensor, which can be used under visible light and can exhibit good PEC performance due to the existence of the heterojunction structure. The experimental results show that the prepared CdS-TiO2NTs electrode had a linear response to 2-16 mM glutathione (GSH). The sensor's sensitivity and detection limit (LOD) were 102.9 µA·mM-1 cm-2 and 27.7 µM, respectively. Moreover, the biosensor had good stability, indicating the potential application of this kind of heterojunction PEC biosensor.

Introducing visible-light sensitivity into photocatalytic CeO2 nanoparticles by hybrid particle preparation exploiting plasmonic properties of gold: enhanced photoelectrocatalysis exemplified for hydrogen peroxide sensing.

In this report we combine the catalytic properties of CeO2 nanoparticles with their transduction ability for photoelectrochemical sensing. This study highlights the usage of CeO2 providing catalytic activity towards H2O2, but only with a limited excitation range in the UV for the construction of a sensing system. In order to improve the photoelectrocatalysis of CeO2 nanoparticles by extending their excitation to visible light, Au/CeO2 core/shell hybrid nanoparticles have been synthesized. The hybrid nanoparticles are fixed on electrodes, allowing for the generation of photocurrents, the direction of which can be controlled by the electrode potential (without bias). The application of the hybrid nanoparticles results in an enhanced photocurrent amplitude under white light illumination as compared to the pure CeO2 nanoparticles. Wavelength-dependent measurements confirm the participation of the Au core in the signal transduction. This can be explained by improved charge carrier generation within the hybrid particles. Thus, by using a plasmonic element the photoelectochemical response of a catalytic nanoparticle (i.e. CeO2) has been spectrally extended. The effect can be exploited for sensorial hydrogen peroxide detection. Here higher photocatalytic current responses have been found for the hybrid particles fixed to gold electrodes although the catalytic reduction has been comparable for both types of nanoparticles. Thus, it can be demonstrated that Au/CeO2 core-shell nanoparticles allow the utilization of visible light for photoelectrochemical hydrogen peroxide (H2O2) detection with improved sensitivity under white light illumination or application of such particles with only visible light excitation, which is not possible for pure CeO2. With help of the layer-by-layer (LbL) technique for nanoparticle immobilization, the electrode response can be adjusted and with a 5 layers electrode a low detection limit of about 3 µM H2O2 with a linear detection range up to 2000 µM is obtained.

Ultrasensitive photoelectrochemical microcystin-LR immunosensor using carboxyl-functionalized graphene oxide enhanced gold nanoclusters for signal amplification.

An ultrasensitive photoelectrochemical (PEC) immunosensor based on gold nanoclusters (AuNCs) with 11-mercaptoundecanoic acid (MUA) ligands was fabricated for determination of microcystin-LR (MC-LR). The PEC immunosensor was developed by loading the monoclonal MC-LR antibody (Ab) to the MUA-AuNCs modified gold electrodes. After different measurement conditions being optimized, silver nanoparticles (AgNPs), gold nanorods (AuNRs), graphene oxide (GO) and carboxyl-functionalized graphene oxide (cGO) were introduced into MUA-AuNCs to enhance the sensing properties. The experimental result revealed that the sensitivity of PEC immunosensors was enhanced by both their photoelectrochemical properties and antibody loading properties with dependent relationship, which was different from the enhancement strategy of PEC sensors based on redox reactions. Among different hybrid nanocomposites, MUA-AuNCs/cGO not only improved the photoelectrochemical properties, but also loaded more antibodies for sensing, which resulted in best sensing performance. Thus, a universal method was proposed to enhance the sensing performance of PEC immunosensors based on impedance changes. Finally, MUA-AuNCs/cGO based PEC immunosensors exhibited a wide linear range of 0.001 nM-1000 nM with low detection limit of 0.011 pM (S/N = 3) for MC-LR determination. Meanwhile, the designed PEC immunosensors showed high selectivity, reproducibility and specificity, which provided the promising applications in aquatic environment.

Albumin Fusion at the N-Terminus or C-Terminus of HM-3 Leads to Improved Pharmacokinetics and Bioactivities.

HM-3, an integrin antagonist, exhibits anti-tumor biological responses and therefore has potential as a therapeutic polypeptide. However, the clinical applications of HM-3 are limited by its short half-life. In this study, we genetically fused human serum albumin (HSA) to the N or C-terminus of HM-3 to improve HM-3 pharmacokinetics. HM-3/HSA proteins were successfully expressed in Pichia pastoris and displayed improved pharmacokinetic properties and stability. Among them, the half-life of HM-3-HSA was longer than HSA-HM-3. In vitro, the IC50 values of HSA-HM-3 and HM-3-HSA were 0.38 ± 0.14 µM and 0.25 ± 0.08 µM in B16F10 cells, respectively. In vivo, the inhibition rates of B16F10 tumor growth were 36% (HSA-HM-3) and 56% (HM-3-HSA), respectively, indicating antitumor activity of HM-3-HSA was higher than HSA-HM-3. In conclusion, these results suggested that the HM-3/HSA fusion protein might be potential candidate HM-3 agent for treatment of melanoma and when HSA was fused at the C-terminus of HM-3, the fusion protein had a higher stability and activity.