Microvascularized tumor assembloids model for drug delivery evaluation in colorectal cancer-derived peritoneal metastasis.

Peritoneal metastasis (PM) is a fatal state of colorectal cancer, and only a few patients may benefit from systemic chemotherapy. Although hyperthermic intraperitoneal chemotherapy (HIPEC) brings hope for affected patients, the drug development and preclinical evaluation of HIPEC are seriously lagging behind, mainly due to the lack of an ideal in vitro PM model that makes drug development over-reliant on expensive and inefficient animal experiments. This study developed an in vitro colorectal cancer PM model [microvascularized tumor assembloids (vTA)] based on an assembly strategy of endothelialized microvessels and tumor spheroids. Our data showed that the in vitro perfusion cultured vTA could maintain a similar gene expression pattern to their parental xenografts. Also, the drug penetration pattern of the in vitro HIPEC in vTA could mimic the drug delivery behavior in tumor nodules during in vivo HIPEC. More importantly, we further confirmed the feasibility of constructing a tumor burden-controlled PM animal model using vTA. In conclusion, we propose a simple and effective strategy to construct physiologically simulated PM models in vitro, thus providing a basis for PM-related drug development and preclinical evaluation of locoregional therapies. STATEMENT OF SIGNIFICANCE: This study developed an in vitro colorectal cancer peritoneal metastasis (PM) model based on microvascularized tumor assembloids (vTA) for drug evaluation. With perfusion culture, vTA could maintain a similar gene expression pattern and tumor heterogeneity to their parental xenografts. And the drug penetration pattern in vTA was similar to the drug delivery behavior in tumor nodules under in vivo treatment. Moreover, vTA was more conducive to construct PM animal models with controllable tumor burden. In conclusion, the construction of vTA could provide a new strategy for the PM-related drug development and preclinical evaluation of locoregional therapies.

Stoma-site approach single-port laparoscopic versus conventional multi-port laparoscopic Miles's procedure for low rectal cancer: A prospective, randomized controlled trial.

OBJECTIVE: The aim of this study was to compare perioperative outcomes of patients with low rectal cancer after stoma-site approach single-port laparoscopic Miles procedure or conventional multi-port laparoscopic Miles procedure, as well as to evaluate the safety and efficacy of stoma-site approach single-port laparoscopic surgery in low rectal cancer. METHODS: Between September 2020 and September 2021, 51 low rectal cancer patients scheduled for Miles procedure at the Department of Gastrointestinal Surgery of Affiliated Hospital of North Sichuan Medical College were randomly assigned to the single-port laparoscopic surgery group (SPLS) and the multi-port laparoscopic surgery (MPLS) group. The perioperative outcomes were compared between the two groups. RESULTS: In this study, 25 patients underwent SPLS and 26 underwent MPLS. All patients completed the study, and there were no perioperative deaths in either group. Observation indicators such as intraoperative bleeding (39 mL vs. 41 mL), number of lymph nodes (20.12 ± 3.29 vs. 21.84 ± 3.74), average hospital stay (7.15 ± 1.52 vs. 7.64 ± 1.66), and time to flatulence (2.5d vs. 2.5d) showed no significant differences between the SPLS and MPLS groups (p > 0.05). However, the operation duration (180 min vs. 118 min) and perioperative complications showed statistically significant differences between the two groups (p < 0.05). In addition, patients in the SPLS group had significantly higher satisfaction scores than those in the MPLS group (p < 0.05). CONCLUSION: For patients with low rectal cancer requiring Miles surgery, stoma-site approach single-port laparoscopic surgery has comparable safety and efficacy to multi-port laparoscopic surgery.

Syntaxin 6 Enhances the Progression of Epithelial Ovarian Cancer by Promoting Cancer Cell Proliferation.

OBJECTIVES: The aim of this study is to evaluate the expression of syntaxin 6 (STX6) in epithelial ovarian cancer (EOC) and assess the effects of STX6 on the prognosis of patient. METHODS: Using information from the Kaplan-Meier Plotter database, the effects of STX6 expression on overall survival (OS) and progression-free survival (PFS) in ovarian cancer patients were examined. The clinical information of 147 patients with epithelial ovarian cancer was evaluated, and immunohistochemical staining was used to identify STX6 expression in postoperative tumor specimens, and the affection of STX6 expression on patient prognosis was assessed. In addition, the expression of STX6 in tumor tissue, peritoneal metastases (PM) derived from 13 patients with epithelial ovarian cancer and 6 normal ovarian specimens was detected by PCR and Western blot. In order to investigate how STX6 affects the proliferation of tumor cells, STX6 was also over expressed and knock down in ovarian cancer cell lines. Then colony formation assay was used to explore the effect of STX6 regulating on cell proliferation. RESULTS: Kaplan-Meier Plotter enrollment data analysis revealed that patients with overexpressed STX6 had substantially worse OS and PFS than individuals with low STX6 expression. Retrospective study revealed a significant (P<0.05) correlation between the STX6 expression and tumor classifications, tumor stage, peritoneal carcinomatosis index (PCI), and PFS survival of patients. Western blot and PCR findings for fresh samples showed that STX6 was overexpressed in both primary lesions and PM nodules of OC. SKOV3 cell proliferation was shown to be dramatically reduced by STX6 knockdown and promoted by STX6 overexpression, according to the in vitro experiments. CONCLUSION: STX6 may increase the progression of epithelial OC by encouraging the proliferation of cancer cells, indicating that STX6 was a viable therapeutic target of epithelial OC.

Bilayer hydrogel dressing with lysozyme-enhanced photothermal therapy for biofilm eradication and accelerated chronic wound repair.

Biofilms are closely associated with the tough healing and dysfunctional inflammation of chronic wounds. Photothermal therapy (PTT) emerged as a suitable alternative which could destroy the structure of biofilms with local physical heat. However, the efficacy of PTT is limited because the excessive hyperthermia could damage surrounding tissues. Besides, the difficult reserve and delivery of photothermal agents makes PTT hard to eradicate biofilms as expectation. Herein, we present a GelMA-EGF/Gelatin-MPDA-LZM bilayer hydrogel dressing to perform lysozyme-enhanced PTT for biofilms eradication and a further acceleration to the repair of chronic wounds. Gelatin was used as inner layer hydrogel to reserve lysozyme (LZM) loaded mesoporous polydopamine (MPDA) (MPDA-LZM) nanoparticles, which could rapidly liquefy while temperature rising so as to achieve a bulk release of nanoparticles. MPDA-LZM nanoparticles serve as photothermal agents with antibacterial capability, could deeply penetrate and destroy biofilms. In addition, the outer layer hydrogel consisted of gelatin methacryloyl (GelMA) and epidermal growth factor (EGF) promoted wound healing and tissue regeneration. It displayed remarkable efficacy on alleviating infection and accelerating wound healing in vivo. Overall, the innovative therapeutic strategy we came up with has significant effect on biofilms eradication and shows promising application in promoting the repair of clinical chronic wounds.

Efficacy and Safety of Bifidobacterium Quadruple Viable Bacteria Combined with Mesalamine against UC Management: A Systematic Review and Meta-Analysis.

Objective: To systematically assess effectiveness and safety of Bifidobacterium quadruple viable bacteria combined with mesalamine against ulcerative colitis (UC) in the Asian population. Methods: An electronic search was conducted in PubMed, Embase, Cochrane Library, CNKI, VIP, and Wanfang databases for a random collection of controlled trials of Bifidobacterium quadruple viable bacteria combined with mesalamine against UC. Following data screening and extraction, a Cochrane risk assessment tool was adopted to evaluate the quality of the included studies, and RevMan 5.3 and Stata/SE 15.1 software were used for meta-analysis. Results: Nineteen articles which enrolled 1,707 subjects were included ultimately in this study. The experimental group performed better than the control group in improving the Mayo score (MD = -1.94, 95% CI = (-2.69, -1.19), P < 0.00001), increasing the total clinical efficiency (OR = 5.10, 95% CI (3.53, 7.38), P < 0.00001), reducing the levels of IL-8 (SMD = -1.79, 95% CI (-2.36, -1.12), P < 0.00001), increasing the levels of IL-4 (SMD = 1.00, 95% CI (0.60, 1.41), P < 0.00001), and reducing the levels of hsCRP (MD = -3.26, 95% CI (-4.28, -2.25), P < 0.00001), TNF-α (MD = -7.11, 95% CI (-9.23, -5.00), P < 0.00001), ox-LDL (MD = -14.46, 95% CI (-17.20, -11.72), P < 0.00001), and LPO (MD = -3.55, 95% CI (-4.70, -2.39), P < 0.0001) as well as increasing SOD level (SMD = 1.68, 95% CI (1.02, 2.35), P < 0.00001), and adverse reactions were substantially less than that of control (OR = 0.43, 95% CI = (0.28, 0.66), P = 0.0001). Conclusion: In conclusion, the current meta-analysis shows that Bifidobacterium quadruple viable bacterium combined with mesalamine has a satisfactory effect in the treatment of UC in China, and its safety is better than that of mesalamine or Bifidobacterium quadruple viable bacteria alone. However, randomized controlled trials with standardized designs and large sample sizes are still needed for further validation.

A MXene-Based Bionic Cascaded-Enzyme Nanoreactor for Tumor Phototherapy/Enzyme Dynamic Therapy and Hypoxia-Activated Chemotherapy.

The enzyme-mediated elevation of reactive oxygen species (ROS) at the tumor sites has become an emerging strategy for regulating intracellular redox status for anticancer treatment. Herein, we proposed a camouflaged bionic cascaded-enzyme nanoreactor based on Ti3C2 nanosheets for combined tumor enzyme dynamic therapy (EDT), phototherapy and deoxygenation-activated chemotherapy. Briefly, glucose oxidase (GOX) and chloroperoxidase (CPO) were chemically conjugated onto Ti3C2 nanosheets, where the deoxygenation-activated drug tirapazamine (TPZ) was also loaded, and the Ti3C2-GOX-CPO/TPZ (TGCT) was embedded into nanosized cancer cell-derived membrane vesicles with high-expressed CD47 (meTGCT). Due to biomimetic membrane camouflage and CD47 overexpression, meTGCT exhibited superior immune escape and homologous targeting capacities, which could effectively enhance the tumor preferential targeting and internalization. Once internalized into tumor cells, the cascade reaction of GOX and CPO could generate HClO for efficient EDT. Simultaneously, additional laser irradiation could accelerate the enzymic-catalytic reaction rate and increase the generation of singlet oxygen (1O2). Furthermore, local hypoxia environment with the oxygen depletion by EDT would activate deoxygenation-sensitive prodrug for additional chemotherapy. Consequently, meTGCT exhibits amplified synergistic therapeutic effects of tumor phototherapy, EDT and chemotherapy for efficient tumor inhibition. This intelligent cascaded-enzyme nanoreactor provides a promising approach to achieve concurrent and significant antitumor therapy.

Gene-engineered exosomes-thermosensitive liposomes hybrid nanovesicles by the blockade of CD47 signal for combined photothermal therapy and cancer immunotherapy.

CD47, overexpressed on kinds of tumor cells, activates a "don't eat me" signal through binding to signal regulatory protein α (SIRPα), leading to immune escape from the mononuclear phagocyte system (MPS). It is also a huge challenge to deliver therapeutic drugs to the tumor sites due to the short retention time in blood, poor targeting of tumor cells and accelerated clearance by MPS. Herein, we designed a hybrid therapeutic nanovesicles, named as hGLV, by fusing gene-engineered exosomes with drug-loaded thermosensitive liposomes. We demonstrated that the CD47-overexpressed hGLV exhibited the long blood circulation and improved the macrophages-mediated the phagocytosis of tumor cells by blocking CD47 signal. Moreover, the resulted hGLV could remarkably target the homologous tumor in mice, achieving the preferential accumulation at the tumor sites. Importantly, hGLV loading the photothermal agent could achieve the excellent photothermal therapy (PTT) under laser irradiation after the intravenous injection, completely eliminating the tumors, leading to immunogenic cell death and generating substantial tumor-associated antigens, which could promote the maturation of immature dendritic cells with the help of the co-encapsulated immune adjuvant to trigger strong immune responses. Generally, the hybrid nanovesicles based on CD47 immune check point blockade can be a promising platform for the drug delivery in cancer treatment.

Thermosensitive Exosome-Liposome Hybrid Nanoparticle-Mediated Chemoimmunotherapy for Improved Treatment of Metastatic Peritoneal Cancer.

Metastatic peritoneal carcinoma (mPC) is a deadly disease without effective treatment. To improve treatment of this disease, a recently developed hyperthermic intraperitoneal chemotherapy (HIPEC) has emerged as the standard of care. However, the efficacy of this approach is limited by inefficient drug penetration and rapidly developed drug resistance. Herein, a nanotechnology approach is reported that is designed to improve drug delivery to mPC and to augment the efficacy of HIPEC through delivery of chemoimmunotherapy. First, the drug delivery efficiency of HIPEC is determined and it is found that chemotherapy agents cannot be efficiently delivered to large tumors nodules. To overcome the delivery hurdle, genetically engineered exosomes-thermosensitive liposomes hybrid NPs, or gETL NPs, are then synthesized, and it is demonstrated that the NPs after intravenous administration efficiently penetrates into mPC tumors and releases payloads at the hypothermia condition of HIPEC. Last, it is shown that, when granulocyte-macrophage colony-stimulating factor (GM-CSF) and docetaxel are co-delivered, gETL NPs effectively inhibit tumor development and the efficacy is enhanced when HIPEC is co-administered. The study provides a strategy to improve drug delivery to mPCs and offers a promising approach to improve treatment of the disease through combination of locoregional delivery of HIPEC and systemic delivery of chemoimmunotherapy via gETL NPs.

Engineered Human Adipose Stem-Cell-Derived Exosomes Loaded with miR-21-5p to Promote Diabetic Cutaneous Wound Healing.

Diabetic wounds are a worldwide health problem causing extremely heavy public health burden and require effective treatment. Optimal strategies for treating nonhealing diabetic wounds include stem-cell-based therapy and delivery of novel drug substances, such as functional microRNAs (miRNAs); however, miRNA easily degrades in the wound microenvironment. Herein, we developed a human adipose stem-cell-derived exosome (hASC-exos)-based miRNA delivery strategy to enhance its therapeutic efficacy. The miR-21-5p mimics, as novel therapeutic candidates for diabetic wounds, were loaded into hASC-exos by electroporation, taking advantage of natural availability and biocompatibility of exosomes as extracellular miRNA transporting particles. The engineered exosomes (E-exos) exhibited excellent effects on promoting proliferation and migration of keratinocytes via Wnt/ß-catenin signaling in vitro and accelerating diabetic wound healing by increasing re-epithelialization, collagen remodeling, angiogenesis, and vessel maturation in vivo. Results from this study would set the fundamentals of applying hASC-exos to deliver future drug substances and to develop cell-free therapy for wound-healing treatments.

Immobilized Candida antarctica lipase B catalyzed synthesis of biodegradable polymers for biomedical applications.

Enzymes, as green catalysts, can catalyze varied reactions with high efficiency. Among various types of enzymes, Candida antarctica lipase B (CALB) is the most commonly used one, due to its high catalytic activity in polymer synthesis. This review highlights the polymers synthesized via the catalysis of immobilized CALB, and focuses on their applications in biomedical areas, especially their use in drug and gene delivery systems. Upholding the idea of sustainable and environment-friendly green chemistry, and emphasizing the safety of products applied for biomedical uses, lipase catalysts are better choices compared to inorganic metal catalysts. We introduced the background and advantages of lipase catalysis and reviewed the lipase-catalyzed synthetic routes involving ring opening polymerization, polycondensation and others. For biomedical applications, we mainly discuss enzymatically synthesized polyesters, due to their outstanding biodegradability and synthetic possibilities. We firstly introduced our early work as basic thinking, then we summarized the subsequent work that focused on developing multiple functional polyesters for different biomedical applications. Prominent experimental results have been shown and the significance of CALB-catalyzed synthesis has been proved.

Downregulation of miR-200a-3p induced by hepatitis B Virus X (HBx) Protein promotes cell proliferation and invasion in HBV-infection-associated hepatocarcinoma.

BACKGROUND: Hepatitis B Virus X (HBx) Protein encoded by HBV is believed to be the major player in the process of HBV-induced oncogenesis. Ectopic expression of miR-200a-3p was reported to be associated with diverse tumorigenesis. This study aimed to better understand the role of miR-200a-3p and its correlation with HBx in HBV-induced hepatocellular carcinoma (HCC). METHODS: In this report, we examined the gene expression using quantitative RT-PCR and protein expression using Western blotting analysis. Cells were transfected with miR-200a-3p mimics or empty vector, and HBx-carrying vector or empty vector. Cell viability was tested using CCK-8 assay. Wound healing assay was performed to assess cell migration while Transwell assay was performed to evaluate cell invasion. RESULTS: miR-200a-3p was downregulated in HBV-positive tissue samples compared with HBV-negative tissue samples. This result was further confirmed with HBV-positive and - negative cell lines. HBx protein was overexpressed in HBV-positive cells where expression of miR-200a-3p was significantly suppressed. Increased cell viability, altered cell cycle progression, increased cell migration and invasion occurred in HBx-overexpressed cells compared to its controls. In forced expressed miR-200a-3p cells, cell viability, cell migration and invasion were significantly decreased, and cell cycle status was altered compared to its controls. CONCLUSIONS: Taken together, pathogenetic function of HBx is negatively correlated with miR-200a-3p in HBV-cased HCC through regulating cell viability, cell cycle arrest, cell migration and cell invasion.