Immunostimulatory gene therapy combined with checkpoint blockade reshapes tumor microenvironment and enhances ovarian cancer immunotherapy

Immune evasion has made ovarian cancer notorious for its refractory features, making the development of immunotherapy highly appealing to ovarian cancer treatment. The immune-stimulating cytokine IL-12 exhibits excellent antitumor activities. However, IL-12 can induce IFN-γ release and subsequently upregulate PDL-1 expression on tumor cells. Therefore, the tumor-targeting folate-modified delivery system F-DPC is constructed for concurrent delivery of IL-12 encoding gene and small molecular PDL-1 inhibitor (iPDL-1) to reduce immune escape and boost anti-tumor immunity. The physicochemical characteristics, gene transfection efficiency of the F-DPC nanoparticles in ovarian cancer cells are analyzed. The immune-modulation effects of combination therapy on different immune cells are also studied. Results show that compared with non-folate-modified vector, folate-modified F-DPC can improve the targeting of ovarian cancer and enhance the transfection efficiency of pIL-12. The underlying anti-tumor mechanisms include the regulation of T cells proliferation and activation, NK activation, macrophage polarization and DC maturation. The F-DPC/pIL-12/iPDL-1 complexes have shown outstanding antitumor effects and low toxicity in peritoneal model of ovarian cancer in mice. Taken together, our work provides new insights into ovarian cancer immunotherapy. Novel F-DPC/pIL-12/iPDL-1 complexes are revealed to exert prominent anti-tumor effect by modulating tumor immune microenvironment and preventing immune escape and might be a promising treatment option for ovarian cancer treatment.

Newly developed gas-assisted sonodynamic therapy in cancer treatment

Sonodynamic therapy (SDT) is an emerging noninvasive treatment modality that utilizes low-frequency and low-intensity ultrasound (US) to trigger sensitizers to kill tumor cells with reactive oxygen species (ROS). Although SDT has attracted much attention for its properties including high tumor specificity and deep tissue penetration, its anticancer efficacy is still far from satisfactory. As a result, new strategies such as gas-assisted therapy have been proposed to further promote the effectiveness of SDT. In this review, the mechanisms of SDT and gas-assisted SDT are first summarized. Then, the applications of gas-assisted SDT for cancer therapy are introduced and categorized by gas types. Next, therapeutic systems for SDT that can realize real-time imaging are further presented. Finally, the challenges and perspectives of gas-assisted SDT for future clinical applications are discussed.

Tumor-targeted/reduction-triggered composite multifunctional nanoparticles for breast cancer chemo-photothermal combinational therapy

Breast cancer has become the most commonly diagnosed cancer type in the world. A combination of chemotherapy and photothermal therapy (PTT) has emerged as a promising strategy for breast cancer therapy. However, the intricacy of precise delivery and the ability to initiate drug release in specific tumor sites remains a challenging puzzle. Therefore, to ensure that the therapeutic agents are synchronously delivered to the tumor site for their synergistic effect, a multifunctional nanoparticle system (PCRHNs) is developed, which is grafted onto the prussian blue nanoparticles (PB NPs) by reduction-responsive camptothecin (CPT) prodrug copolymer, and then modified with tumor-targeting peptide cyclo(Asp-d-Phe-Lys-Arg-Gly) (cRGD) and hyaluronic acid (HA). PCRHNs exhibited nano-sized structure with good monodispersity, high load efficiency of CPT, triggered CPT release in response to reduction environment, and excellent photothermal conversion under laser irradiation. Furthermore, PCRHNs can act as a photoacoustic imaging contrast agent-guided PTT. In vivo studies indicate that PCRHNs exhibited excellent biocompatibility, prolonged blood circulation, enhanced tumor accumulation, allow tumor-specific chemo-photothermal therapy to achieve synergistic antitumor effects with reduced systemic toxicity. Moreover, hyperthermia-induced upregulation of heat shock protein 70 in the tumor cells could be inhibited by CPT. Collectively, PCRHNs may be a promising therapeutic way for breast cancer therapy.

Intracellular aggregation of peptide-reprogrammed small molecule nanoassemblies enhances cancer chemotherapy and combinatorial immunotherapy

The intracellular retention of nanotherapeutics is essential for their therapeutic activity. The immobilization of nanotherapeutics inside target cell types can regulate various cell behaviors. However, strategies for the intracellular immobilization of nanoparticles are limited. Herein, a cisplatin prodrug was synthesized and utilized as a glutathione (GSH)-activated linker to induce aggregation of the cisplatin prodrug/IR820/docetaxel nanoassembly. The nanoassembly has been reprogrammed with peptide-containing moieties for tumor-targeting and PD-1/PD-L1 blockade. The aggregation of the nanoassemblies is dependent on GSH concentration. Evaluations

Can nanoparticles and nano‒protein interactions bring a bright future for insulin delivery?

Insulin therapy plays an essential role in the treatment of diabetes mellitus. However, frequent injections required to effectively control the glycemic levels lead to substantial inconvenience and low patient compliance. In order to improve insulin delivery, many efforts have been made, such as developing the nanoparticles (NPs)-based release systems and oral insulin. Although some improvements have been achieved, the ultimate results are still unsatisfying and none of insulin-loaded NPs systems have been approved for clinical use so far. Recently, nano‒protein interactions and protein corona formation have drawn much attention due to their negative influence on the

Correlation between cardiac resynchronization response and pulmonary artery hemodynamic parameters / 中南大学学报(医学版)

OBJECTIVES@#To evaluate the response to cardiac resynchronization therapy (CRT) and the correlation between CRT and pulmonary artery hemodynamic parameters.@*METHODS@#The patients with chronic heart failure indicator for CRT were enrolled. The left ventricular end-systolic volume (LVESV) was measured by echocardiography and New York Heart Association (NYHA) classification was evaluated between one week before and six months after CRT. Mean pulmonary artery pressure (mPAP), pulmonary artery systolic pressure (PASP) and pulmonary vascular resistance (PVR) were measured by right heart catheterization. Left ventricular reverse remodeling (LVRR) is defined as a decrease of 15% or more in LVESV at the 6th month after CRT; Clinical response is defined as a decrease of NYHA classification at or above grade 1 at the 6th month after CRT. Pulmonary hypertension (PH) was defined as mPAP≥25 mmHg. According to the response, patients were divided into 3 groups: group A (LVRR+clinical response), group B (no LVRR+clinical response) and group C (no LVRR+no clinical response). The changes of NYHA classification, echocardiographic and pulmonary hemodynamic parameters were observed in the 3 groups. The Kaplan-Meier survival curve was used to analyze the differences in all-cause mortality, combined end-point events of death or re-hospitalization due to heart failure among different groups.@*RESULTS@#A total of 45 patients with CRT implantation [aged (63.27±9.55) years, 36 males] were included. The average follow-up period was (33.76±11.50) months. Thirty-one patients (68.89%) were in group A, 9 of whom with PH. Eight patients (17.78%) were in group B, 7 of whom with PH. Six patients were in group C, all with PH. Cardiac function including NYHA classification, echocardiographic and pulmonary hemodynamic parameters had been significantly improved in group A after CRT implantation (0.05). There were no significant changes in NYHA classification, echocardiographic and pulmonary hemodynamic parameters in group C (>0.05). Compared with group C, group A and group B had lower all-cause mortality (=0.005) and lower incidence of composite endpoint events (=0.001).@*CONCLUSIONS@#Patients with LVRR and clinical response after CRT have a good prognosis. Patients with clinical response but without LVRR have a better prognosis than those without clinical response and LVRR, which may be related to the decrease of pulmonary hemodynamic parameters such as mPAP and TPG.

Reproductive and developmental toxicity of 2- (2H-1, 2, 3-benzotriazol-2-yl) -4-methyl-6- (2-methylpropen-2-yl) phenol in mice: an experimental study / 中华劳动卫生职业病杂志

Objective@#To investigate the reproductive and developmental toxicity of 2- (2H-1, 2, 3-benzotriazol-2-yl) -4-methyl-6- (2-methylpropen-2-yl) phenol in mice and to provide a basis for its risk assessment.@*Methods@#The reproductive and developmental toxicity of 2- (2H-1, 2, 3-benzotriazol-2-yl) -4-methyl-6- (2-methylpropen-2-yl) phenol was tested using the screening method of chemicals with reproductive and developmental toxicity in "Chemical Testing Method" (SEPA). After five days of adaptive feeding, 120 specific pathogen-free healthy Kunming mice (male/female ratio=1:1) were orally administered 0 (control) , 146, 292, and 584 mg/kg 2- (2H-1, 2, 3-benzotriazol-2-yl) -4-methyl-6- (2-methylpropen-2-yl) phenol for two weeks. One male mouse was mated with one female mouse in a single cage. The day on which a vaginal plug was observed was defined as gestation day 0 (GD0). The exposure for female mice was sustained to four days postpartum and the exposure for male mice was sustained for two weeks after mating. The body weight, food intake, body length, tail length, and sex ratio were recorded and the reproductive index was calculated. The reproductive organs were weighed and subjected to histopathological examination.@*Results@#The 584 mg/kg group had significantly lower body weight at weeks 5 and 6 and food intake at week 6 in male mice, uterus weight and uterus/body weight ratio in female mice, and body weight, body length, and tail length on day 0 in offspring compared with the control group (all P<0.05). The 292 mg/kg group had significantly lower testis weight of male mice and food intake of female mice at gestational week 2 than the control group (both P<0.05). The 146 mg/kg group had significantly lower food intake of female mice at gestational week 2 than the control group (P<0.05) .@*Conclusion@#For male and female Kunming mice, the no observed adverse effect levels of 2- (2H-1, 2, 3, -benzotriazol-2-yl) -4-methyl-6- (2-methylpropen-2-yl) phenol are both 146 mg/kg.

Gold nanorods together with HSP inhibitor-VER-155008 micelles for colon cancer mild-temperature photothermal therapy

Enhancing the heat-sensitivity of tumor cells provides an alternative solution to maintaining the therapeutic outcome of photothermal therapy (PTT). In this study, we constructed a therapeutic system, which was composed of methoxy-polyethylene-glycol-coated-gold-nanorods (MPEG-AuNR) and VER-155008-micelles, to evaluate the effect of VER-155008 on the sensitivity of tumor cells to heat, and further investigate the therapeutic outcome of MPEG-AuNR mediated PTT combined with VER-155008- micelles. VER-155008- micelles down-regulate the expression of heat shock proteins and attenuate the heat-resistance of tumor cell. The survival of HCT116 cells treated with VER-155008- micelles under 45 °C is equal to that treated with high temperature hyperthermia (55 °C) . Furthermore, we proved either the MPEG-AuNR or VER-155008- micelles can be accumulate in the tumor site by photoacoustic imaging and fluorescent imaging. anti-cancer evaluation showed that tumor size remarkably decreased (smaller than 100 mm or vanished) when treated with combing 45 °C mild PTT system, which contrasted to the tumor size when treated with individual 45 °C mild PTT (around 500 nm) or normal saline as control (larger than 2000 nm). These results proved that the VER-155008- micelles can attenuate the heat-resistance of tumor cells and enhance the therapeutic outcome of mild-temperature photothermal therapy.

Effects of PRX-2 gene on the phenotype changes of epidermal stem cells differentiating into sweat gland cells / 中华整形外科杂志

Objective@#To investigate the effects of PRX-2 gene on phenotype changes in epidermal stem cells differentiating into sweat gland cells.@*Methods@#Epidermal stem cells and sweat gland cells separated and cultured from healthy foreskin and adult full-thick skin respectively, were identified by immunofluorescence staining. Lentiviral vector-mediated overexpression and knockdown of PRX-2 gene in epidermal stem cells were performed respectively, with empty vector-mediated epidermal stem cells as a control group. Overexpression、blank control and knowdown group′s PRX-2 expressions in gene and protein levels were detected using RT-PCR and Western blot technology. The ESCs of each group were co-cultured with sweat gland cells through transwell plate, and the expressions of CEA and β1 integrin in epidermal stem cells were determined by flow cytometry before and after co-culturing.@*Results@#Epidermal stem cells and sweat gland cells were in line with their respective specific antigens. Before co-cultured, epidermal stem cells highly expressed β1 integrin (98.69±0.67)%, hardly expressed CEA (6.20±3.15)%. After co-cultured, β1 integrin expression levels were showed as knockdown group (19.30±0.53)%<blank control group (65.77±2.32)% <overexpress group (92.63±10.97)%, and CEA expression levels as knockdown (95.43±2.36)%> blank control group (51.20±0.79)%> overexpress group (45.91±0.93)%. There had significant differences between those of each two groups.@*Conclusions@#PRX-2 gene can inhibit the phenotypic change of Epidermal Stem Cells differentiating into Sweat Gland Cells and improve the ability to maintain their own specific antigens.

Construction and identification of recombinant lentivirus carrying human beta defensins-3 and connective tissue growth factor gene / 军事医学

Objective To construct a recombinant lentivirus containing human beta defensins -3 ( hBD3 ) , connective tissue growth factor gene (CTGF) and enhanced green fluorescent protein (EGFP), and to detect its translation in rabbit bone marrow mesenchymal stem cells (BMSC).Methods The lentivirus containing hBD3, CTGF and EGFP genes was constructed in vitro.The titer of lentivirus was tested with end-paint dilution assay .Rabbit BMSCs were transfected with recombinant virus.The best value of multiplicity of infection (MOI) was tested.The expression condition, transfection efficacy and genetic stability of the target genes were evaluated by using fluorescence microscopy and flow cytometry . Western blotting was used to detect the expression of the target protein .Results Recombinant lentivirus vectors: Lenti-CTGF-hBD3-EGFP, Lenti-hBD3-EGFP, and Lenti-EGFP, were successfully obtained . The titer of the recombinant lentiviruses was 3.21 ×108, 5.80 ×108, and 1.16 ×109, respectively.The best MOI value to transfect BMSCs was 150. The transfection efficacy of these lentivirus vectors was high , reaching 79.72%as assessed by flow cytometry , and it could be stably inherited .Western blotting displayed that target protein expression was successful .Conclusion The construction of recombinant lentiviruses carrying hBD3 and CTGF genes is successful and can be effectively transfected into BMSCs .

Anticoagulant chitosan nanoparticles：synthesis, characterization and biological safety / 中国组织工程研究

BACKGROUND:Studies have shown that chitosan and other natural polysaccharides have heparin-like anticoagulant function after sulfonated modification. Sulfonated chitosan has good anticoagulant property because the sulfonate group formed by sulfonated chitosan is similar with the active group of heparin. OBJECTIVE: To prepare the anticoagulant chitosan nanoparticles and to detect its morphology, physical and chemical properties and biological security. METHODS: Chitosan nanoparticles were synthesized by emulsion-chemical cross link. Sulfonated chitosan nanoparticles were synthesized by sulfonation reaction. Its morphology was described by transmission electron microscope. The peak-value change of its specific groups was observed by infrared spectroscopy. (1) Coagulation experiment: Heparin, chitosan nanoparticles and 10, 30 and 50 mg of sulfonated chitosan nanoparticles were added into the blood of Spraque-Dawley rats. The coagulation indicators were detected. (2) Hemolysis experiment: deionized water, physiological saline and 10, 30, 50 g/L sulfonated chitosan nanoparticles extracts were added into 2% red blood cel suspension of rabbits. The hemolysis rate was detected. (3) Cytotoxicity experiments: DMEM medium containing fetal bovine serum and 10, 30, 50 g/L sulfonated chitosan nanoparticle extracts were used to culture human umbilical vein endothelial cels. Cel relative growth rate and toxicity grading were detected after 72 hours. RESULTS AND CONCLUSION: Scanning electron microscopy showed that sulfonated chitosan nanoparticles had good morphology, with a diameter of 50 nm. Infrared spectroscopy showed that the sulfonated replacement occurred.In vitro coagulation experiments showed that sulfonated chitosan nanoparticles had significant anticoagulant effects in a dose-dependent manner. Sulfonated chitosan nanoparticles meet the national safety standard for hemolysis rate of less than 5%, non-induced hemolysis property. Cytotoxicity assays showed that sulfonated chitosan nanoparticles extracts had no significant cytotoxicity, and its biological safety was in line with the national standards.

Intelligent Stretching and Its Application in Spasticity and Contracture of Ankle Joint (review) / 中国康复理论与实践

@#Contracture and spasticity of ankle joints were major sources of disability in neurological impairment including stroke and cerebral palsy, etc. The manual stretching used in physical therapy might be laborious and time-consuming to the therapists and the outcome was dependent on the experience and the subjectiveend feelingof the therapists. A device was developed that could safely stretch the an-kle joint to its extreme positions with quantitative control of the resistance torque and stretching velocity. Furthermore, it could satisfy a strong need for quantitative and objective measures of the impairment and rehabilitation outcome. This was just the meaning intelligent stretching referred to. This article described the origin of the concept of intelligent stretching and its definition, operational principle, and su-periority and weakness, as well as its application in ankle joint spasticity and contracture in patients with stroke and cerebral palsy.

Current progress in research on the nanomaterials for biological macromolecules recognition / 生物医学工程学杂志

The combination of nanotechnology and molecular imprinting technique, including their application research in biomedical domain, provides a new solution to the problem of the substitutes for antibodies, enzymes, and other native biological structures as well as cell bracket materials. Nanocavity biomaterials with recognition specificity imprinted by using proteins as templates have numerous applications in biotechnology, medicine and so on. This review presents the aspects of molecular imprinting nanostructure involved in the biomacromolecules imprinting, and it explores the precent developments and achievements of nanomaterials for molecular imprinting technology.

A novel nano-hydroxyapatite/aliphatic polyesteramide composite / 生物医学工程学杂志

Hydrothermally synthesized nano-hydroxyapatite (n-HA) varying in weight from 10% to 30% was used as filler to make nanocomposites with novel aliphatic polyesteramide (PEA) in our laboratory. The structure and properties of PEA and its n-HA composites were investigated through transmission electron microscopy, infrared spectrometry, X-ray diffractioin, scanning electron microscopy and energy spectrometry. The shape and size of the n-HA crystals are similar to those of the apatite crystals in natural bone. Molecule interactions are present between the n-HA and PEA in the composite, which allows the uniform dispersion of n-HA in PEA matrix. This contributes enhanced mechanical property and bioactivity to the composite. The cytocompatibility of the composites has been investigated by culturing osteoblasts on the membranes. Good cell attachment and proliferation manner were observed on the membranes after 1 week. These results suggest that the PEA/n-HA composites prepared in this study may serve as potential candidate scaffold for tissue engineering.

Synthesis and characterization of thermosensitive hydrogel based on PEG-PCL-PEG block copolymers / 生物医学工程学杂志

In this paper, a series of low-molecular-weight PEG-PCL-PEG triblock copolymers were successfully synthesized by ring-opening polymerization method, and were characterized using 1H-NMR and FTIR. The aqueous solution displayed specific thermosensitive gel-sol transition when the concentration was above corresponding critical gel concentration (CGC). The gel-sol phase diagram was recorded using test tube-inverting method, which was depended on the hydrophilic/hydrophobic balance in macromolecular structure, as well as heating history. As a result, the gel-sol transition temperature range could be altered, which might be very useful for its application as injectable drug delivery system.

Effects of Volatile Organic Compounds Escaped from Alkyd Dope on Neurotransmitter Content in Brain of Mice / 环境与健康杂志

Objective To investigate the effects of volatile organic compounds escaped from alkyd dope on neurotransmitter content in the brain of mice.Methods 56 BALB/c mice were randomly divided into 7 groups,8 in each and exposed to volatile organic compounds(30 mg/m3)escaped from alkyd dope by dynamic inhalation exposure.The neurotransmitters(cholines,monoamines,amino acids)in the brain were determined during the period of exposure and comeback.Results Compared with the control,DA,5-HT,NE and Glu significantly decreased and Gly,GABA significantly increased during the period of exposure,however NE still kept significant decrease during the period of comeback.Conclusion Volatile organic compounds escaped from alkyd dope may cause neurochemical change in mice.