Dual-Engineered Macrophage-Microbe Encapsulation for Metastasis Immunotherapy.

Lung metastases are the leading cause of death among cancer patients. The challenges of inefficient drug delivery, compounded by a robust immunosuppressive microenvironment, make effective treatment difficult. Here, an innovative dual-engineered macrophage-microbe encapsulation (Du-EMME) therapy is developed that integrates modified macrophages and engineered antitumor bacteria. These engineered macrophages, termed R-GEM cells, are designed to express RGD peptides on extracellular membranes, enhancing their tumor cell binding and intratumor enrichment. R-GEM cells are cocultured with attenuated Salmonella typhimurium VNP20009, producing macrophage-microbe encapsulation (R-GEM/VNP cells). The intracellular bacteria maintain bioactivity for more than 24 h, and the bacteria released from R-GEM/VNP cells within the tumor continue to exert bacteria-mediated antitumor effects. This is further supported by macrophage-based chemotaxis and camouflage, which enhance the intratumoral enrichment and biocompatibility of the bacteria. Additionally, R-GEM cells loaded with IFNγ-secreting strains (VNP-IFNγ) form R-GEM/VNP-IFNγ cells. Treatment with these cells effectively halts lung metastatic tumor progression in three mouse models (breast cancer, melanoma, and colorectal cancer). R-GEM/VNP-IFNγ cells vigorously activate the tumor microenvironment, suppressing tumor-promoting M2-type macrophages, MDSCs, and Tregs, and enhancing tumor-antagonizing M1-type macrophages, mature DCs, and Teffs. Du-EMME therapy offers a promising strategy for targeted and enhanced antitumor immunity in treating cancer metastases.

Programmable Bacteria with Dynamic Virulence Modulation System for Precision Antitumor Immunity.

Engineered bacteria-mediated antitumor approaches have been proposed as promising immunotherapies for cancer. However, the off-target bacterial toxicity narrows the therapeutic window. Living microbes will benefit from their controllable immunogenicity within tumors for safer antitumor applications. In this study, a genetically encoded microbial activation strategy is reported that uses tunable and dynamic expression of surface extracellular polysaccharides to improve bacterial biocompatibility while retaining therapeutic efficacy. Based on screening of genes associated with Salmonella survival in macrophages, a novel attenuated Salmonella chassis strain AIS (htrA gene-deficient) highly enriched in tumors after administration and rapidly cleared from normal organs are reported. Subsequently, an engineered bacterial strain, AISI-H, is constructed based on the AIS strain and an optimized quorum-sensing regulatory system. The AISI-H strain can achieve recovery of dynamic tumor-specific bacterial virulence through a novel HTRA-RCSA axis-based and quorum-sensing synthetic gene circuit-mediated increase in extracellular polysaccharide content. These strains act "off" in normal organs to avoid unwanted immune activation and "on" in tumors for precise tumor suppression in mice. The AISI-H strain shows significant tumor inhibition and potent activation of anticancer immunity in a melanoma mouse model. The AISI-H strain exhibits excellent biocompatibility. This bacterial regulation strategy expands the applications of microbe-based antitumor therapeutics.

Development and Validation of the Coronary Heart Disease Scale Among the System of Quality of Life Instruments for Chronic Diseases QLICD-CHD (V2.0) Based on Classical Test Theory and Generalizability Theory.

Objective: Coronary heart disease (CHD) is a common and frequent disease with a long and incurable course, and the quality of life of patients is severely reduced. This study was to develop and validate a quality of life scale for patients with CHD based on the Chinese context. Methods: The scale QLICD-CHD (V2.0) was developed based on the QLICD-CHD (V1.0), using a programmed decision procedures. Based on the data measuring QoL 3 times before and after treatments from 189 patients with CHD, the psychometric properties of the scale were evaluated with respect to validity, reliability and responsiveness employing correlation analysis, multi-trait scaling analysis, structural equation modeling, t-test and also G-study and D-study of generalizability theory analysis. The SF-36 scale was used as the criterion to evaluate the criterion-related validity. Paired t tests were conducted to evaluate the responsiveness on each domain/facet as well as the total of the scale, with Standardized Response Mean (SRM) being calculated. Results: The QLICD-CHD (V2.0) has been developed with 42 items in 4 domains. The Cronbach's α of the general module, the specific module and the total scale were 0.91, 0.92 and 0.91 respectively. The overall score and the test-retest reliability coefficients in all domains are higher than 0.60, except for the specific module. Correlation and factor analysis confirmed good construct validity and criterion-related validity. After treatments, the overall score and score of all domains have statistically significant changes (P<0.01). The SRM of domain-level score ranges from 0.27 to 0.50. Generalizability Theory further confirm the reliability of the scale through more accurate variance component studies. Conclusion: The QLICD-CHD (V2.0) could be used as a useful instrument in assessing QoL for patients with CHD, with good psychometric properties.

Camouflaging attenuated Salmonella by cryo-shocked macrophages for tumor-targeted therapy.

Live bacteria-mediated antitumor therapies mark a pivotal point in cancer immunotherapy. However, the difficulty in reconciling the safety and efficacy of bacterial therapies has limited their application. Improving bacterial tumor-targeted delivery while maintaining biosafety is a critical hurdle for the clinical translation of live microbial therapy for cancer. Here, we developed "dead" yet "functional" Salmonella-loaded macrophages using liquid nitrogen cold shock of an attenuated Salmonella typhimurium VNP20009-contained macrophage cell line. The obtained "dead" macrophages achieve an average loading of approximately 257 live bacteria per 100 cells. The engineered cells maintain an intact cellular structure but lose their original pathogenicity, while intracellular bacteria retain their original biological activity and are delay freed, followed by proliferation. This "Trojan horse"-like bacterial camouflage strategy avoids bacterial immunogenicity-induced neutrophil recruitment and activation in peripheral blood, reduces the clearance of bacteria by neutrophils and enhances bacterial tumor enrichment efficiently after systemic administration. Furthermore, this strategy also strongly activated the tumor microenvironment, including increasing antitumor effector cells (including M1-like macrophages and CD8+ Teffs) and decreasing protumor effector cells (including M2-like macrophages and CD4+ Tregs), and ultimately improved antitumor efficacy in a subcutaneous H22 tumor-bearing mouse model. The cryo-shocked macrophage-mediated bacterial delivery strategy holds promise for expanding the therapeutic applications of living bacteria for cancer.

New method for large-scale facial skin sebum quantification and skin type classification.

BACKGROUND: The demand for personalized skin care is increasing rapidly in recent years. In order to provide suitable products for different customers, an accurate assessment of the skin condition is required. It is necessary to establish a remote and convenient method for quantifying skin condition. MATERIALS AND METHODS: We established a new method for quantifying the facial sebum excretion, using Sebutape as well as a model for objective skin type classification. RESULTS: In total, 2173 sebum quantification samples were collected and categorized into 4 skin types. We found good agreement between subjective and objective skin types. 60.33% of the detection is consistent, and the proportion of discrepancies is only 0.55%. The main mismatch is caused by broad criteria for combination type classification. Quantifying the sebum excretion will help to describe and analyze the skin state easily. We found sebum levels in Chinese women peak at the age between 20 and 30 and decrease dramatically after age 40s. CONCLUSION: This new sebum quantitative method is well consistent to self-report skin type. In addition, it provides a powerful tool for the cosmetic industry to evaluate skin condition precisely and recommend personalized products accordingly.

A new method for skin aging evaluation of Chinese women.

BACKGROUND: With the development of cosmetic industry in China, an aging evaluation method of Chinese women is in great need. AIMS: The aim of this study is to establish a skin aging assessment method using convenient portable machine Antera 3D. METHODS: The quantitative approach was developed by capturing 11 areas of the face, and overall, 48 parameters were extracted for aging evaluation. Later, 297 subjects were recruited to take facial image and life style questionnaire. Evaluation of age was accomplished by establishing prediction method with detected skin traits. RESULTS: The age prediction model was built by using the evaluated facial traits, and a R square of 0.6 is achieved by comparing to the chronological age. The crow's feet, cheek skin pigmentation, forehead skin tone, and wrinkles around the month are critical factors in evaluating Chinese women skin aging. In addition, we also explored life styles associated with important skin aging traits. CONCLUSION: The method developed in this research provides reliable alternative in aging study of Chinese women.

Nationwide Survey and Identification of Potential Stress Factor in Sensitive Skin of Chinese Women.

PURPOSE: Sensitive skin is characterized by uncomfortable sensations in response to a number of factors. We performed a large-scale study to investigate the prevalence of sensitive skin at all ages and the impacts of related factors across China. METHODS: A nationwide sampling of the Chinese population aged over 18 was conducted. Subjects were categorized into sensitive and non-sensitive groups, and critical differences between these two groups were investigated. RESULTS: In total, 22,085 questionnaires were collected from Chinese women with sensitive skin. The prevalence of sensitive skin is 49.6% and is associated with age, skin type, geographic area of subjects, and other factors. Heavy life stress and the application of several cosmetic products also affect the prevalence of sensitive skin. CONCLUSION: Having a combination or oily skin type, living in the municipalities, being under heavy stress, and applying several cosmetic products will increase skin stress and contribute to the occurrence of sensitive skin.

RhBMP-2 immobilized on poly(phthalazinone ether nitrile ketone) via chemical and physical modification for promoting in vitro osteogenic differentiation.

Poly(ether ether ketone) (PEEK) is a polyaryletherketone commonly used for bone implants, but it is difficult to modify the PEEK surface. Conversely, poly(phthalazinone ether nitrile ketone) (PPENK) is a polyaryletherketone whose surface can be modified by using chemical reactions owing to its cyano group. In this paper, two types of materials, P-BMP-2 and PH-BMP-2, were prepared by covalent immobilization and heparin binding of rhBMP-2 respectively to enhance the osteogenic activity of PPENK. X-ray photoelectron spectroscopy and water contact-angle measurement were used to demonstrate the hydrolysis of the cyano groups on PPENK, amine group grafting and immobilization of rhBMP-2. Immunohistochemical staining and evaluation of loading and release behaviour were used to demonstrate the existence of rhBMP-2 on PPENK surfaces. The biological activity of MC3T3-E1 preosteoblast cells on the samples were evaluated using cell adhesion, viability and proliferation tests. The genetic expression of genes associated with osteogenic activity was assessed by reverse transcription polymerase chain reaction. Based on the obtained in vitro experimental results, both P-BMP-2 and PH-BMP-2 exhibit good cytocompatibility and promote differentiation of MC3T3-E1 preosteoblast cells. In particular, the favourable biocompatibility can be obtained using the heparin-binding method.