A healthy live birth after mosaic blastocyst transfer in preimplantation genetic testing for GATA1-related cytopenia combined with HLA matching.

BACKGROUND: GATA1-related cytopenia (GRC) is characterized by thrombocytopaenia and/or anaemia ranging from mild to severe. Haematopoietic stem cell transplantation (HSCT) is a healing therapeutic choice for GRC patients. We identified a novel pathogenic variant (GATA1: c.1019delG) in a boy with GATA1-related cytopenia. Then we performed preimplantation genetic testing (PGT) in this GRC family. After a mosaic embryo transfered, a healthy and HLA-compatible with the proband baby was delivered. CASE PRESENTATION: The proband is a 6-year-old boy who was diagnosed to have transfusion-dependent anaemia since 3 year old. Whole-exome sequencing (WES) showed that the proband has a hemizygous variant c.1019delG in GATA1, which is inherited from his mother. His parents decided to undergo PGT to have a health and HLA-compatible offspring. After whole genome amplification (WGA) of biopsied trophectoderm (TE) cells, next generation sequencing (NGS)-based PGT was preformed to analyse embryos on chromosomal aneuploidy, target mutation and HLA typing. There were 3 embryos HLA-matched to the proband. The genotypes of the 3 embryos were heterozygous variant, hemizygous variant, normal respectively. After a heterozygous, mosaic partial trisomy (chr)16, and HLA-matched embryo transfer, a healthy baby was delivered and whose HSCT is compatible with the proband. CONCLUSIONS: NGS-based PGT-HLA is a valuable procedure for the treatment of GATA1-related cytopenia caused by GATA1 variants, or other haematological disorders, oncological and immunological diseases. Furthermore, our study reconfirms that mosaic embryos transfer would bring healthy offspring.

Selection of functional sperm by using hyaluronic acid modified magnetic microbeads and an electromagnetic manipulation system.

In recent years, the incidence of infertility has increased year by year. Assisted reproductive technology (ART) is one of the effective strategies to treat infertility. In the process of ART, commonly used methods for sperm separation have shortcomings and there is still room for improvement. In this study, a functional sperm selection strategy was established based on hyaluronic acid (HA) modified magnetic microbeads (MBs) and a supporting two-dimensional electromagnetic manipulation device system. Due to the modification of HA on the surface of MBs, the HA-MBs have the ability to target and bind to specific receptors on the sperm membrane to form a sperm-MB complex. A disulfide linker was introduced to connect HA and MBs. After modifying HA and connecting it with the disulfide linker, the sperm of sperm-MBs can be released under the combined effect of hyaluronidase and reduced GSH with the disulfide bond broken and HA degraded. A two-dimensional electromagnetic manipulation system was introduced to generate a magnetic field and control the directional movement of the sperm-MB complex under the guidance of an inverted microscope. The free MBs can also be removed after the sperm is released. Furthermore, the mouse sperm selected through this strategy can achieve normal insemination via ICSI and the obtained blastocysts have normal morphology and developmental milestones. This strategy has potential to be developed into an automated screening solution for the screening of functional sperm for assisted reproductive technology.

MiR-140-3p Normalizes Vascular Smooth Muscle Cell Behavior by Inhibiting the RhoA/ROCK Signaling Pathway in Lower-extremity Arteriosclerosis Obliterans.

BACKGROUND: Excessive vascular smooth muscle cell (VSMC) proliferation and migration are the main contributors to the symptoms of lower-extremity arteriosclerosis obliterans (ASO). Previous studies suggested that microRNAs (miRNAs) regulate VSMC activity. Nevertheless, the molecular mechanisms by which they do so are unclear. OBJECTIVE: The present study aimed to identify the biological processes accounting for the effects of miR-140-3p on VSMCs in ASO. METHODS: The expression levels of miR-140-3p in clinical samples were analyzed by real-time polymerase chain reaction. An ASO cell model was established to investigate the expression of miR-140-3p on VSMCs. The transwell® assays and MTT assays were used to assess migration and proliferation. The interaction between RhoA and miR-140-3p was verified using the Dualluciferase reporter assay. Western blot technique was used to identify RhoA, RhoA-associated protein kinase 1 (ROCK1), and ROCK2. RESULTS: We discovered that miR-140-3p inhibited the proliferation, migration, and invasion but promoted the apoptosis of VSMCs, and RhoA was its downstream target gene. RhoA, ROCK1, and ROCK2 were upregulated in vascular tissues damaged by ASO compared to normal, healthy arteries. MiR-140-3p also decreased RhoA, ROCK1, and ROCK2 mRNA and protein expression. CONCLUSION: Overall, the present work partially elucidated the mechanism by which miR-140-3p regulates VSMC function and offered novel insights into potential therapeutic approaches for patients with lower-extremity arteriosclerosis obliterans.

Three-dimensional treatment-planning-based prediction of seed migration to chest after 125I seed brachytherapy for hepatic malignancy.

PURPOSE: To develop and validate risk models incorporating clinical and/or imaging parameters based on three-dimensional treatment-planning systems (3D-TPS) to predict the occurrence of 125I seed migration and the number of migrated seeds <2/≥2 to the chest after brachytherapy for patients with malignant hepatic tumors. METHODS AND MATERIALS: A total of 480 patients diagnosed with malignant liver tumors receiving 125I seed brachytherapy from July 2010 to May 2020 were retrospectively enrolled. Variables included 3D-TPS-based CT parameters, that is, the distance from the seed to the inferior vena cava (DSI), the distance from the seed to the second hepatic portal (DSP) and the angle from the seed to the second hepatic portal (ASP), and patients' clinical characteristics, that is, the number of seed implantation procedures (NSP), the maximum number of implanted seeds one time (MAX) and laboratory parameters within 1 week before treatment. Two sets of logistic regression models incorporating clinical and/or imaging variables were developed to predict the occurrence of seed migration and the number of migrated seeds <2/≥2. Model performance was assessed by ROC analysis and decision curve analysis. RESULTS: Compared with the clinical models, the combined model showed a higher discriminative ability for both the prediction of migration occurrence and number of migrated seeds ≥ 2/<2 to the chest (AUC, 0.879 vs. 0.668, p < 0.05; 0.895 vs. 0.701, p < 0.05). The decision curve analysis results indicated higher net benefits of combined models than clinical models. Variables, including DSI, NSP and pretreatment lymphocyte-to-neutrophil ratio, acted as the most important predictors in combined models. CONCLUSIONS: The proposed combined models based on 3D-TPS improved discriminative abilities for predicting 125I seed migration and number of migrated seeds <2/≥2 to the chest after hepatic brachytherapy, being promising to aid clinical decision-making.

First preimplantation genetic testing case of Meckel syndrome with a novel homozygous TXNDC15 variant in a non-consanguineous Chinese family.

BACKGROUND: Meckel-Gruber syndrome (MKS) is a perinatally lethal, genetically heterogeneous, autosomal recessive condition caused by defective primary cilium formation. So far, the association of TXNDC15-related MKS has been reported in only five independent families from diverse ethnic origins, including Saudi, Pakistani, Estonian, and Indian. Here, we report a fetus diagnosed with MKS at 12 weeks, exhibiting typical ultrasound findings. METHODS: Low-coverage whole-genome sequencing was used to identify chromosomal abnormalities. Trio-base whole exome sequencing (trio-WES) was performed to investigate the potential pathogenic variants associated with MKS. Preimplantation genetic testing for monogenic disorders (PGT-M) was applied to prevent the transmission of the pathogenic variant. RESULTS: A novel homozygous pathogenic variant in the TXNDC15 gene was identified through trio-WES. The application of PGT-M successfully prevented the transmission of the pathogenic variant and resulted in an ongoing pregnancy. CONCLUSION: This is the first report of a TXNDC15 variant in the Chinese population and the first PGT case of TXNDC15-related MKS worldwide. The successful application of PGT-M in this family provides a potential approach for other monogenic diseases. Our case expands the variant spectrum of TXNDC15 and contributes to the molecular diagnosis and genetic counseling for MKS. This case underscores the importance of appropriate genetic testing methods and accurate genetic counseling in the diagnosis of rare monogenic diseases.

Increased Expression of CD74 in Atherosclerosis Associated with Inflammatory Responses of Endothelial Cells and Macrophages.

To clarify the relationship between CD74 and atherosclerosis (AS) and the mechanisms in oxidized LDL (ox-LDL)-induced endothelial cell and macrophage injury. Datasets obtained from the Gene Expression Omnibus database are integrated. Differentially expressed genes (DEGs) were obtained using R software. Weighted gene co-expression network analysis (WGCNA) was performed to screen the target genes. The endothelial cell injury model and macrophage foaming model were established using ox-LDL, and CD74 expression was detected by Quantitative reverse transcription PCR (RT-qPCR) and Western blot (WB). Then, after silencing CD74, cell viability and ROS production were measured, and WB detected the expression of p-p38 MAPK and NF-κB. There were 268 DEGs associated with AS, of which CD74 was up-regulated. The turquoise module containing CD74 in WGCNA was positively associated with AS. Cell viability was significantly decreased in the endothelial cell injury and macrophage foaming models, while CD74, ROS production, NF-κB, and p-p38MAPK expression increased (P < 0.05). After silencing CD74, ROS production, NF-κB, and p-p38MAPK expression were decreased and cell viability was higher than the model group (P < 0.05). CD74 is up-regulated in endothelial cell injury and macrophage foaming models and is involved in AS progression via the NF-κB and MAPK signaling pathways.

Safety and efficacy of biliary stenting combined with iodine-125 seed strand followed by hepatic artery infusion chemotherapy plus lenvatinib with PD-1 inhibitor for the treatment of extrahepatic cholangiocarcinoma with malignant obstructive jaundice.

Objectives: To evaluate the efficacy and safety of biliary stenting implantation with iodine-125 seed strand (SI) followed by hepatic artery infusion chemotherapy (HAIC) plus lenvatinib (Len) with programmed death-1 (PD-1) inhibitor for patients diagnosed with extrahepatic cholangiocarcinoma (ECC) and malignant obstructive jaundice (MOJ). Methods: In this single-center retrospective study, the data of ECC patients with MOJ from March 2015 to January 2023 was assessed. Using probability score matching (PSM), the selection bias of patients was reduced. Primary study outcomes included overall survival (OS) and progression-free survival (PFS). The OS and PFS were performed using the Kaplan-Meier method and evaluated with the log-rank test. Results: A total of 104 patients were enrolled finally, including 52 patients treated with interventional therapy (SI+HAIC) plus Len with PD-1 inhibitor (SI+HAIC+Len+P group) and 52 patients treated with interventional therapy (SI+HAIC) plus lenvatinib (SI+HAIC+Len group). 26 pairs of patients were matched after PSM analysis. After PSM analysis, the median OS and PFS in the SI+HAIC+Len+P group were significantly longer compared to those in the SI+HAIC+Len group (OS:16.6 vs. 12.3 months, P = 0.001; PFS:12.6 vs 8.5 months, P = 0.004). The DCR was significantly different between groups (P = 0.039), while ORR not (P = 0.548). The addition of PD-1 inhibitor was generally well tolerated without treatment-associated mortality. Conclusion: Interventional therapy (SI+HAIC) plus Len with PD-1 inhibitor was effective for ECC patients accompanied by MOJ with a manageable safety profile.

A meta-analysis comparing hepatic arterial infusion chemotherapy and sorafenib for advanced hepatocellular carcinoma.

Background: The majority of patients with hepatocellular carcinoma (HCC) are diagnosed in an advanced stage. Although sorafenib is recommended as the standard treatment for advanced HCC, its efficacy is limited. In some studies, hepatic arterial infusion chemotherapy has demonstrated a significant therapeutic benefit for advanced HCC compared with sorafenib. We systematically evaluated and compared the efficacy and safety of hepatic arterial infusion chemotherapy and sorafenib for advanced HCC. Methods: A systematic search of PubMed, Embase, Web of Science and the Cochrane Library up to December 31, 2020 was conducted. Study outcomes included overall survival (OS), progression-free survival (PFS), objective response rate (ORR), disease control rate (DCR), and adverse effects. The hazard ratio and odds ratio with 95% confidence intervals (CI) were used to measure the pooled effect. Results: Eighteen retrospective or prospective cohort studies and one prospective controlled study were included, with 1,339 patients treated with hepatic arterial infusion chemotherapy (HAIC) and 1,060 patients treated with sorafenib. We found that hepatic arterial infusion chemotherapy was superior to sorafenib in terms of OS [hazard ratio (HR): 0.66, 95% CI: 0.46-0.95, P=0.027], PFS (HR: 0.55, 95% CI: 0.44-0.69, P<0.001), ORR [assessed using Response Evaluation Criteria in Solid Tumors (RECIST): OR: 9.02, 95% CI: 6.01-13.53, P<0.001; assessed using modified RECIST: odds ratio (OR): 3.71, 95% CI: 1.92-7.16, P<0.001], and DCR (assessed using RECIST: OR: 2.31, 95% CI: 1.40-3.83, P=0.001; assessed by modified RECIST: OR: 2.28, 95% CI: 1.22-4.28, P=0.01). Dermatological adverse effects and hypertension were significantly higher in the sorafenib group for all grades of adverse effects. However, regarding severe adverse effects, hepatic arterial infusion chemotherapy was associated with more frequent leukocytopenia and thrombocytopenia. Conclusions: Hepatic arterial infusion chemotherapy demonstrated favorable efficacy and safety for advanced HCC compared with sorafenib and should be recommended for suitable patients with advanced HCC.

Constitutive TDO2 expression promotes liver cancer progression by an autocrine IL-6 signaling pathway.

BACKGROUND: Increased tryptophan (Trp) metabolism by indoleamine 2,3-dioxygenase (IDO)/tryptophan 2,3-dioxygenase (TDO) represents one of the most studied pathways for immunosuppression in tumor tissues. However, the pro-tumor effects induced by Trp metabolism remain controversial. METHODS: The paraffin sections of tumor tissues were obtained from patients with liver cancer and examined by immunohistochemical staining to investigate the role of Trp metabolic enzymes. To further confirm the pro-tumor effects induced by TDO2, we established TDO2 overexpression SMC-7721 and HepG2 liver cancer cell lines, and western blotting, cell proliferation, and colony formation were evaluated. Meanwhile, liver cancer subcutaneous mice models were established, and the tumorigenic rates of SMC-7721 cells, tumor volume and survival of bearing mice were calculated. In addition, the survival data of liver cancer patients from The Cancer Genome Atlas (TCGA) database were downloaded to analyze the effect of TDO2 expression on the survival of patients with liver cancer. RESULTS: Here, we showed that constitutive TDO2 expression gave rise to liver cancer through upregulation of Trp metabolism. And the TDO2 expression was positively correlated with the poor prognosis in liver cancer patients. TDO2 expression in tumor cells accounted for the release of kynurenine (Kyn), which activated aryl hydrocarbon receptor (AhR) to promote liver cancer cells proliferation. Mechanistically, we found that AhR expression contributed to the secretion of Interleukin-6 (IL-6), thereby promoting tumor cells proliferation through the STAT3 and NF-kB/TIM4 signals. Interrupt of AhR signals by PDM2 revealed improved outcomes in subcutaneous tumor-bearing mice. CONCLUSIONS: Together, our study showed that the TDO2/Kyn/AhR/IL-6 signaling pathway was a novel mechanism underlying the malignancy of liver cancer, and suggested that AhR signals might be a valuable therapeutic target for tumor therapy.

Evaluation of percutaneous transcatheter embolization for pulmonary arteriovenous malformations.

BACKGROUND: The purpose of this study was to assess the safety and efficacy of percutaneous transcatheter embolization (TCE) for the treatment of pulmonary arteriovenous malformations (PAVMs). METHODS: Forty-three consecutive patients (n = 17 males; n = 26 females) with 72 untreated PAVMs underwent coil and/or plug embolization between January 2010 and February 2018. The mean patient age was 42 ± 14 years (range 19-71 years). The median size of the feeding artery was 7.9 ± 2.9 mm (range 3.5-14.0 mm). The arterial blood gas level and cardiac function of all patients were analysed. The technical success rate, recanalization rate, and complications were evaluated. Computed tomography angiography (CTA) examinations were scheduled for 12 months after treatment and every 2-4 years thereafter. RESULTS: Twenty-five PAVMs were treated with coils alone, twenty-one were treated with plugs alone, and twenty-six were treated with both coils and plugs. The technical success rate was 100%. There were no complications during operation. However, one patient (2.3%) had pulmonary thrombosis and embolism post-operation. The patients' pre-operative and post-operative PaO2 and SaO2 levels were significantly different (p < 0.01). A comparison of the New York Heart Association (NYHA) grade before and after embolization in all patients showed a significant decrease in the post-operative grade (p < 0.01). The 72 PAVMs were divided into three groups (coils only group [n = 25], plugs only group [n = 21], and coils/plugs combined group [n = 26]). After 12 months of follow-up, there were seven reperfusion PAVMs in the coil group, seven reperfusion PAVMs in the plug group, and 1 reperfusion PAVM in the combined group. There were significant differences between the two groups and the combined group. CONCLUSION: Percutaneous TCE is safe and effective for the treatment of PAVMs. A combination of coils and vascular plugs may be useful for preventing recanalization after the embolization of PAVMs.

Using copper sulfide nanoparticles as cross-linkers of tumor microenvironment responsive polymer micelles for cancer synergistic photo-chemotherapy.

Photo-chemotherapy presents promising therapeutic effects in cancer treatment. Photo-thermal and chemotherapeutic agents are generally delivered independently or jointly by drug carriers, such as polymer micelles. A polymer micelle is one type of widely researched drug carrier. However, there is a disassembly risk for polymer micelles under excessive dilution in blood circulation, leading to the premature release of payloads from the micelles and finally resulting in undesirable toxic side effects. Herein, amino-PEG decorated copper sulfide nanoparticles (CuS NPs) with photothermal effect were applied as a cross-linker to enhance polymeric micelles' stability and to provide photothermal therapy in the meanwhile. The micelles were prepared using a pH/reductive responsive polymer, poly(ε-caprolactone)-ss-poly(2-(diisopropylamino)ethyl methacrylate/glycidyl methacrylate/2-methylacrylloxyethyl phosphorylcholine (PCL-SS-P(DPA/GMA/MP)), abbreviated as DGM. Cross-linked micelles (DGM-CuS) exhibited high photothermal transformation efficiency and excellent stability against dilution, as well as pH and redox responsive drug release. Under near-infrared laser irradiation, the cell cytotoxicity of doxorubicin-loaded micelles DGM-CuS@DOX and DGM-CuS@DOX-P (DGM-CuS@DOX modified by peptides) increased by 17.1 times and 69.2 times correspondingly compared to that without laser irradiation. All of the solid 4T1 tumors disappeared, and tumor metastases were merely observed in the major organs of the tumor-bearing mice after administration of DGM-CuS@DOX and DGM-CuS@DOX-P with irradiation. In this synergistic therapy system, CuS NPs play double roles of a photothermal agent and a micelle cross-linker. The strategy of utilizing nanoparticles as cross-linkers is newly reported, which offers new insight for combination therapy.

Effect of surface morphology change of polystyrene microspheres through etching on protein corona and phagocytic uptake.

Surface physicochemistry properties of polymer particles are crucial for protein corona formation and macrophage phagocytosis when they contact with living body. In this work, polystyrene microspheres (PS-MSs) were selected as a model of polymer microparticles and fabricated by chromic acid etching through controlling conditions to obtain different surface morphology structures and to investigate their effect on the protein adsorption and phagocytic uptake of PS-MSs. The adsorption of bovine serum albumin (BSA) and fibrinogen (FIB) on PS-MSs showed almost the same tendency, i.e. the etched PS-MSs presented lower protein adsorption compared with original microspheres. The adsorption of BSA and FIB was the lowest when the protuberances on the etched surfaces were maximum and the size of the protuberances was minimum. Furthermore, the surface morphologies of PS-MSs were influenced in return not only by the amounts of proteins but also by protein types. Meanwhile, the macrophages phagocytosis of PS-MSs depended on the amounts and kinds of adsorbed proteins, especially the albumin content. In a word, phagocytosis and protein adsorption can be regulated by microsphere morphologies through etching, which provides a promising strategy to avoid invalid uptake for polymer particles such as drug delivery carriers.

Exogenous vitamin C triggered structural changes of redox-activated dual core-crosslinked biodegradable nanogels for boosting the antitumor efficiency.

Premature leakage of drugs during blood circulation and slow drug release at the tumor site are two major challenges that nanocarriers have to overcome to achieve successful cancer therapy. Herein, a dual core-crosslinked, redox-sensitive polymeric nanogel (sDL) was constructed by the self-assembly of two star-shaped amphiphilic copolymers (4sP(EG-b-LLA)-N3, 4sP(EG-b-DLA)-N3) in the presence of a redox-sensitive crosslinker (d-ss-Bu), where hydrophilic polyethylene glycol (PEG) was used as the shell and the functional hydrophobic poly(l-lactide) (PLLA) and poly(d-lactide) (PDLA) were used as the dual crosslinked core via stereocomplex formation and chemical interactions. The dual core-crosslinked structure of the nanogels allowed for almost 2-fold enhanced doxorubicin (DOX)-loading capacity, favorable structural stability to restrict the premature leakage of therapeutic drug and smaller particle size to accelerate the internalization efficiency compared to non-crosslinked nanocarriers. Furthermore, exogenous vitamin C (Vc) can trigger the breakage of redox-sensitive bonds to accelerate drug release from nanogels for improved in vitro antitumor efficacy. Notably, in vivo near-infrared imaging showed that the highly stable DOX-loaded sDL efficiently aggregated at the tumor site. Sequential administration of DOX-loaded sDL and Vc exhibited the highest tumor inhibition effect without associated systemic toxicity compared to the corresponding single injection of Vc or DOX-loaded sDL control groups for in vivo studies, indicating that exogenous administration of Vc can synergistically impact the release of DOX from sDL. Therefore, the developed nanogels proved to be promising smart carriers for achieving precise tunable-stability in response to relevant environments and the combination of Vc to activate reduction-sensitive drug delivery is a promising approach to maximize the therapeutic efficacy.

Synergistic action of doxorubicin and 7-Ethyl-10-hydroxycamptothecin polyphosphorylcholine polymer prodrug.

There are opportunities for improvements to the efficiency and toxicity of widely used cancer chemotherapy agents such as doxorubicin (DOX·HCl) and 7-Ethyl-10-hydroxycamptothecin (SN38). We developed a safe and effective combination therapy by encapsulating DOX into micelles of a zwitterionic polymer prodrug, SN38 conjugate of poly (α-azide caprolactone-co-caprolactone)-b-poly (2-methacryloyloxyethyl phosphorylcholine [P(CL/CL-g-SN38)-b-PMPC)] which was described in our previous work. The polymer prodrug micelles displayed a higher loading capacity of DOX due to the π-π stacking effect between DOX and SN38 in comparison with the micelles self-assembled by prodrug's precursor, poly(α-azide caprolactone-co-caprolactone)-b-poly(2-methacryloyloxyethyl phosphorylcholine (P(ACL/CL)-b-PMPC). The DOX loaded prodrug micelles decelerated the release of DOX, and also prolonged its circulation. The micelles showed favorable cellular internalization by 4T1 cells. DOX loaded SN38 prodrug micelles displayed good in vitro anticancer effects owing to the synergistic action of doxorubicin and SN38 and were as effective as DOX·HCl, but with lower toxicity than DOX·HCl. Given the synergetic effects of free drug and polymer prodrug, this nanomedicine may offer a safe and effective drug delivery methodology for conventional drug formations.

Exogenous Vitamin C-Triggered Surface Charge Conversion of pH/Reduction-Responsive Micelles for the Enhanced Tumor-Specific Activity of Loaded Doxorubicin.

The effective accumulation at tumor sites and endocytosis by tumor cells for anticancer agents in carriers are essential in successful cancer therapy, and both of the processes are affected by the surface charge of drug carriers. In this study, vitamin C (VC) was employed as an "exogenous switch" to trigger the surface charge conversion of DOX-loaded micelles to obtain a better antitumor effect. T micelles formed by poly(ε-caprolactone)-b-poly(N,N-diethylaminoethyl methacrylate)-ss-b-poly(2-methacryloyloxyethyl phosphorylcholine) (PCL-PDEA-ss-PMPC) turned their ζ potentials from +1 mV to +18 mV under treatment of 20 mM VC, while the ζ potentials of control R micelles formed by PCL-ss-P(DEA-r-MPC) almost remained unchanged under the same condition. DOX-loaded T@DOX and R@DOX had high DLCs of 12% and 13.8%, respectively, and both showed an accelerated drug release in a reductive environment (10 mM GSH or 20 mM VC) at pH 5.0. Notably, due to the surface charge conversion and fast drug release triggered by VC, T@DOX/VC (T@DOX was pretreated by VC) showed an enhanced cytotoxicity and cellular uptake superior to T@DOX, R@DOX, and R@DOX/VC. T@DOX/VC also displayed the in vivo antitumor effect well, which was comparable to DOX·HCl but with less toxic side effects than DOX·HCl. In summary, VC as an exogenous trigger can induce a better antitumor effect of drug-loaded micelles with a suitable polymer structure by charge conversion, and T@DOX/VC has shown to be as a promising approach to achieve potent treatment of tumors.

pH/redox dual-responsive amphiphilic zwitterionic polymers with a precisely controlled structure as anti-cancer drug carriers.

Responding to the tumor microenvironment, functional polymers can serve as preeminent drug carriers for targeted cancer therapy. Stimuli-responsive polymeric drug carriers are reported with diverse anti-tumor effects for various polymer structures. Thus, three pH/redox dual-responsive amphiphilic zwitterionic polymer 'isomers' with different locations of pH/redox responsive units were prepared to understand the relationship between polymer structure and anti-tumor effect. Containing poly(ε-caprolactone) (PCL), poly(N,N-diethylaminoethyl methacrylate) (PDEA) and poly(2-methacryloyloxyethyl phosphorylcholine) (PMPC), polymers PCL-ss-P(DEA-r-MPC) (SDRM), PCL-ss-PDEA-b-PMPC (SDBM) and PCL-PDEA-ss-PMPC (DSM) with a precisely controlled structure were constructed and confirmed through NMR, FITR and EA. The formed micellar drug carriers were characterized by their morphology, loading capacity, acid/redox sensitivity, drug release, in vitro cytotoxicity and in vivo antitumor effects. Micelles with uniform spherical morphologies can effectively encapsulate anti-tumor drugs such as DOX. Among these micelles, DSM@DOX displays the most excellent drug encapsulation capacity (13.4%) with neutral surface charge (-1.02 mV) and good stability, and is different from SDRM@DOX with positive charge (+11.1 mV) and SDBM@DOX with poor stability. All micelles respond to acid and reducing environments and present fast drug release at mildly acidic pH and high concentrations of GSH, exhibiting low burst release under the physiological conditions of plasma. There is no significant difference between these micelles in tumor cell cytotoxicity against MCF-7 and 4T1 cells. Internalization of SDRM@DOX and DSM@DOX by the tumor cells is stronger than that of SDBM@DOX. Notably, DSM@DOX has longer blood circulation and more effective accumulation at the tumor site than the other two micelles. As a result, DSM@DOX shows enhanced antitumor efficacy in 4T1 tumor-bearing mice with reduced side toxicities. Overall, structures of the above polymers significantly influence the in vivo antitumor effects of the drug carriers through blood circulation and cellular uptake.

Effects of nitric oxide donor N,N'-di-sec-butyl-N,N'-dinitroso-1,4-phenylenediamine on the expression of interferon-gamma in tumor infiltrating lymphocytes.

Nitric oxide (NO) has been proven to be a key regulator in the mammalian immune response, such as the innate and adaptive immune responses to tumors. The messenger NO involves T helper cell differentiation and lymphocyte biofunctions. In this study, we employed N,N'-di-sec-butyl-N,N'-dinitroso-1,4-phenylenediamine as NO donor and released NO around tumor infiltrating lymphocytes in vitro by short-time blue light irradiation. The interferon-γ secretion of tumor infiltrating lymphocytes was investigated to study the functional changes caused by the accurate spatio-temporal delivery of NO. The downregulation of interferon-γ in tumor infiltrating lymphocytes after NO treatment indicates promising biological applications to potentially play a role in the treatment of autoimmune diseases. The study was approved by the Medical Ethics Committee of the Shenzhen Second People's Hospital, the First Affiliated Hospital of Shenzhen University, China (approved No. 065) on February 12, 2018.

Bioinspired mimics: Self-assembly of redox-activated phosphorylcholine-based biodegradable copolymers for enhancing antitumor efficiency.

With the purpose of reducing side effects in anticancer therapy, the micelles of a novel reduction-activated copolymer with biomimicking phosphorylcholine, poly(ε-caprolactone)-ss-b-poly(2-methacryloyloxyethyl phosphorylcholine) (PCL-ss-PMPC) are developed. The well-suitable nanosize of micelles with good physiological stability (approximately 50 nm, 2.5 µg/mL) can be quickly internalized into cells due to bioinspired phosphorylcholine property and mainly located in endo/lysosomes. The reduction response of micelles is confirmed by size change and accelerated drug release under reducing environment, proved as better anticancer efficacy in comparison to insensitive micelles. Pharmacokinetics and in vivo studies demonstrate that redox-activated polymeric micelles can prolong blood transportation, facilitate passive target and accumulate in tumor site, and prompt drug release in cytoplasmic redox environment, behaving as much better antitumor efficiency than control and positive DOX·HCl groups. More importantly, the DOX-loaded micelles considerably reduce side effects and systematic toxicity. Therefore, this work fabricated an innovative bioinspired nanosystem via a facile strategy to achieve effective anticancer therapy.

Protein adsorption and macrophage uptake of zwitterionic sulfobetaine containing micelles.

Micelles of poly(ε-caprolactone)-b-poly(N,N-diethylaminoethyl methacrylate)/(N-(3-sulfopropyl-N-methacryloxyethy-N,N-diethylammonium betaine)) (PCL-PDEAPS) and poly(ε-caprolactone)-b-poly(ethylene glycol) (PCL-PEG) were prepared and characterized. The interactions of micelles with model proteins such as bovine serum albumin (BSA), lysozyme (Ly), fibrinogen (Fg) and plasma were studied from adsorption quantity, micelle size, polydispersity index (PDI) and zeta-potential measurements. The adsorption quantity of negatively charged proteins on PCL-PDEAPS micelles containing zwitterionic sulfobetaine is larger than on non-ionic PEG-PCL micelles. The adsorption amount increases with the increase of zwitterionic content. And the quantity of adsorbed Fg is much higher than that of BSA because the former is much larger than the latter. In contrast, adsorption of positively charged Ly on copolymer micelle is very low. The interactions between micelles and model proteins are not only dependent on the hydration of zwitterions in PCL-PDEAPS micelles, but also on the electrostatic effect between proteins and micelles. Moreover, the adsorption of three model proteins on the mixed micelles of PCL-PDEAPS and PCL-PEG copolymers is related to the ratio of two copolymers. Denaturation of the proteins is not detected during adsorption and detachment process. PCL-PDEAPS micelles with positive charge are not swallowed by the macrophages after plasma absorption, in contrast to PCL-PEG micelles.

In vitro and in vivo anti-tumor efficiency comparison of phosphorylcholine micelles with PEG micelles.

Polymer micelles for anticancer drug delivery have shown many advantages. In this study, poly(ε-caprolactone)-b-poly(2-methacryloyloxyethyl phosphorylcholine) (PCL-PMPC) with bio-inspired structure self-assembled into small and uniform micelles as traditional poly(ε-caprolactone)-b-poly(ethylene glycol) (PCL-PEG). The in vitro and in vivo anti-tumor efficiency of PCL-PMPC and PCL-PEG micelles were detailedly evaluated. The both micelles were able to load DOX with high efficiency. PCL-PMPC micelles exhibited faster drug release at pH 5.5 than that of PCL-PEG micelles. Confocal laser scanning microscopy and flow cytometry results showed that PCL-PMPC micelles were more effectively internalized by tumor cells. DOX-loaded PCL-PMPC micelles presented higher cytotoxicity to tumor cells. PCL-PMPC micelles displayed not only longer circulation time in pharmacokinetics investigation, but also higher accumulation at the tumor site in in vivo imaging study in comparison with PCL-PEG micelles. More importantly, in a tumor model DOX-loaded PCL-PMPC micelles showed better therapeutic efficacy than DOX-loaded PCL-PEG micelles along with mild side effects. Therefore, PCL-PMPC micelles are deemed to be promising drug carriers for cancer therapy.