NLRC5 negatively regulates the NF-kappaB and type I interferon signaling pathways.

Stringent control of the NF-kappaB and type I interferon signaling pathways is critical to effective host immune responses, yet the molecular mechanisms that negatively regulate these pathways are poorly understood. Here, we show that NLRC5, a member of the highly conserved NOD-like protein family, can inhibit the IKK complex and RIG-I/MDA5 function. NLRC5 inhibited NF-kappaB-dependent responses by interacting with IKKalpha and IKKbeta and blocking their phosphorylation. It also interacted with RIG-I and MDA5, but not with MAVS, to inhibit RLR-mediated type I interferon responses. Consistent with these observations, NLRC5-specific siRNA knockdown not only enhanced the activation of NF-kappaB and its responsive genes, TNF-alpha and IL-6, but also promoted type I interferon signaling and antiviral immunity. Our findings identify NLRC5 as a negative regulator that blocks two central components of the NF-kappaB and type I interferon signaling pathways and suggest an important role for NLRC5 in homeostatic control of innate immunity.

Delivery of mRNA vaccine with a lipid-like material potentiates antitumor efficacy through Toll-like receptor 4 signaling.

Intracellular delivery of messenger RNA (mRNA)-based cancer vaccine has shown great potential to elicit antitumor immunity. To achieve robust antitumor efficacy, mRNA encoding tumor antigens needs to be efficiently delivered and translated in dendritic cells with concurrent innate immune stimulation to promote antigen presentation. Here, by screening a group of cationic lipid-like materials, we developed a minimalist nanovaccine with C1 lipid nanoparticle (LNP) that could efficiently deliver mRNA in antigen presenting cells with simultaneous Toll-like receptor 4 (TLR4) activation and induced robust T cell activation. The C1 nanovaccine entered cells via phagocytosis and showed efficient mRNA-encoded antigen expression and presentation. Furthermore, the C1 lipid nanoparticle itself induced the expression of inflammatory cytokines such as IL-12 via stimulating TLR4 signal pathway in dendritic cells. Importantly, the C1 mRNA nanovaccine exhibited significant antitumor efficacy in both tumor prevention and therapeutic vaccine settings. Overall, our work presents a C1 LNP-based mRNA cancer nanovaccine with efficient antigen expression as well as self-adjuvant property, which may provide a platform for developing cancer immunotherapy for a wide range of tumor types.

Correction: Gao et al. A Modeling Method for Thermal Error Prediction of CNC Machine Equipment Based on Sparrow Search Algorithm and Long Short-Term Memory Neural Network. Sensors 2023, 23, 3600.

In the original publication [...].

Emerging biomolecules for practical theranostics of liver hepatocellular carcinoma.

Most cases of hepatocellular carcinoma (HCC) are able to be diagnosed through regular surveillance in an identifiable patient population with chronic hepatitis B or cirrhosis. Nevertheless, 50% of global cases might present incidentally owing to symptomatic advanced-stage HCC after worsening of liver dysfunction. A systematic search based on PUBMED was performed to identify relevant outcomes, covering newer surveillance modalities including secretory proteins, DNA methylation, miRNAs, and genome sequencing analysis which proposed molecular expression signatures as ideal tools in the early-stage HCC detection. In the face of low accuracy without harmonization on the analytical approaches and data interpretation for liquid biopsy, a more accurate incidence of HCC will be unveiled by using deep machine learning system and multiplex immunohistochemistry analysis. A combination of molecular-secretory biomarkers, high-definition imaging and bedside clinical indexes in a surveillance setting offers a comprehensive range of HCC potential indicators. In addition, the sequential use of numerous lines of systemic anti-HCC therapies will simultaneously benefit more patients in survival. This review provides an overview on the most recent developments in HCC theranostic platform.

A Modeling Method for Thermal Error Prediction of CNC Machine Equipment Based on Sparrow Search Algorithm and Long Short-Term Memory Neural Network.

To better solve the problem of thermal error of computerized numerical control machining equipment (CNCME), a thermal error prediction model based on the sparrow search algorithm and long short-term memory neural network (SSA-LSTMNN) is proposed. Firstly, the Fuzzy C-means clustering algorithm (FCMCA) is used to screen the key temperature-sensitive points of the CNCME. Secondly, by taking the temperature rise data of key temperature-sensitive points as input and the corresponding time thermal error data as output, we established the SSA-LSTMNN thermal error prediction model. The SSA is used to optimize the parameters of LSTMNN and make its performance play the best. Taking the VMC1060 vertical machining center as the research object, we carried out the experiment. Finally, the prediction effect of the proposed model is compared with the article swarm optimized algorithm and LSTM neural network (PSOA-LSTMNN), the LSTMNN, and the traditional recurrent neural network (TRNN) model. The results show that the average values of the predicted residual fluctuations of the SSA-LSTMNN model are all more than 44% lower than those of the other three models under different operating conditions, which has a strong practicality.

Dynamics of microbial community structure and enzyme activities during the solid-state fermentation of Forgood Daqu: a starter of Chinese strong flavour Baijiu.

Daqu is the traditional fermentation starter for Chinese Baijiu, a traditional Chinese distilled liquor. Although the microbes in Daqu blocks play important roles in the solid-state fermentation process, the changes in microbial community structure and the correlation between the microbiota and enzyme activity have seldom been discussed in previous studies. In this research, a high-throughput Illumina MiSeq sequencing method was used to detect the compositions and changes in microbial diversity in Daqu blocks. The results showed that high-temperature solid fermentation directly changed the main microorganisms from Saccharomycopsis, Wickerhamomyces, Bacillus and Staphylococcus to Aspergillus, Thermoascus, Thermoactinomyces and an unspecified Thermoactinomycetaceae. The richness and diversity of both fungi and bacteria showed a tendency to first increase and then decrease. Through redundancy analysis, it was found that there were positive correlations between certain enzyme activities and certain microbes. (1) Glucoamylase and esterase activities correlated with abundances of Leuconostoc, Weissella, an unspecified Aspergillaceae, an unspecified Trichosporonaceae and an unspecified Ascomycota. (2) Amylase activity correlated with abundances of an unspecified Thermoactinomycetaceae, Thermoactinomyces, Aspergillus and Rasamsonia. (3) Protease activity correlated with abundances of Bacillus, an unspecified Lactobacillus and Saccharomycopsis. In summary, the results of this research provide useful information for understanding and controlling the maturation process of Daqu.

Blocking MCT4 SUMOylation inhibits the growth of breast cancer cells.

Monocarboxylate transporter 4 (MCT4) is highly expressed in various types of solid neoplasms including breast cancer (BC); however, the pro-tumor functions underlying its increased expression have not been explained. Here, we examined the roles of posttranslational modifications to MCT4 in BC, particularly SUMOylation. Our findings revealed that SUMOylation of MCT4 inhibited its degradation and stabilized MCT4 protein levels, while ubiquitination facilitated MCT4 degradation. The E3 ubiquitin ligases ß-TRCP and FBW7 interacted with MCT4 at the DSG-box and TPETS sequences, respectively, and Lys448 (K448) of MCT4 could be modified by SUMO chains. Our key finding was that K448 was crucial for MCT4 SUMOylation. Moreover, mutations of K448 abolished MCT4 expression, delaying the growth of BC. This study suggested that SUMOylation of K448 increased MCT4 levels, and mutations of K448 in MCT4 could have therapeutic significance in BC.

TAK1 negatively regulates NF-κB and p38 MAP kinase activation in Gr-1+CD11b+ neutrophils.

Stringent control of NF-κB and mitogen-activated protein kinase (MAPK) signaling is critical during innate immune responses. TGF-ß activated kinase-1 (TAK1) is essential for NF-κB activation in T and B cells but has precisely the opposite activity in myeloid cells. Specific deletion of TAK1 (Map3k7(ΔM/ΔM)) led to development of splenomegaly and lymphomegaly associated with neutrophilia. Compared with wild-type cells, TAK1-deficient neutrophils enhanced the phosphorylation of the kinases IKK, p38, and JNK and the production of interleukin-1ß (IL-1ß), IL-6, tumor necrosis factor-α (TNF-α), and reactive oxygen species (ROS) after lipopolysaccharide (LPS) stimulation. Map3k7(ΔM/ΔM) mice were significantly more susceptible to LPS-induced septic shock and produced higher amounts of IL-1ß, IL-6, and TNF-α in plasma than do wild-type mice. Specific ablation of p38 rescued the phenotype and functional properties of Map3k7(ΔM/ΔM) mice. Our findings identify a previously unrecognized role of TAK1 as a negative regulator of p38 and IKK activation in a cell type-specific manner.

xCT: A Critical Molecule That Links Cancer Metabolism to Redox Signaling.

System xc- cystine/glutamate antiporter, composed of a light-chain subunit (xCT, SLC7A11) and a heavy-chain subunit (CD98hc, SLC3A2), is mainly responsible for the cellular uptake of cystine in exchange for intracellular glutamate. In recent years, the xCT molecule has been found to play an important role in tumor growth, progression, metastasis, and multidrug resistance in various types of cancer. Interestingly, xCT also exhibits an essential function in regulating tumor-associated ferroptosis. Despite significant progress in targeting the system xc- transporter in cancer treatment, the underlying mechanisms still remain elusive. It is also unclear why solid tumors are more sensitive to xCT inhibitors such as sulfasalazine, as compared to hematological malignancies. This review mainly focuses on the role of xCT cystine/glutamate transporter in regard to tumor growth, chemoresistance, tumor-selective ferroptosis, and the mechanisms regulating xCT gene expression. The potential therapeutic implications of targeting the system xc- and its combination with chemotherapeutic agents or immunotherapy to suppress tumor growth and overcome drug resistance are also discussed.

IKKα inactivation promotes Kras-initiated lung adenocarcinoma development through disrupting major redox regulatory pathways.

Lung adenocarcinoma (ADC) and squamous cell carcinoma (SCC) are two distinct and predominant types of human lung cancer. IκB kinase α (IKKα) has been shown to suppress lung SCC development, but its role in ADC is unknown. We found inactivating mutations and homologous or hemizygous deletions in the CHUK locus, which encodes IKKα, in human lung ADCs. The CHUK deletions significantly reduced the survival time of patients with lung ADCs harboring KRAS mutations. In mice, lung-specific Ikkα ablation (IkkαΔLu ) induces spontaneous ADCs and promotes KrasG12D-initiated ADC development, accompanied by increased cell proliferation, decreased cell senescence, and reactive oxygen species (ROS) accumulation. IKKα deletion up-regulates NOX2 and down-regulates NRF2, leading to ROS accumulation and blockade of cell senescence induction, which together accelerate ADC development. Pharmacologic inhibition of NADPH oxidase or ROS impairs KrasG12D-mediated ADC development in IkkαΔLu mice. Therefore, IKKα modulates lung ADC development by controlling redox regulatory pathways. This study demonstrates that IKKα functions as a suppressor of lung ADC in human and mice through a unique mechanism that regulates tumor cell-associated ROS metabolism.

USP19 modulates autophagy and antiviral immune responses by deubiquitinating Beclin-1.

Autophagy, mediated by a number of autophagy-related (ATG) proteins, plays an important role in the bulk degradation of cellular constituents. Beclin-1 (also known as Atg6 in yeast) is a core protein essential for autophagic initiation and other biological processes. The activity of Beclin-1 is tightly regulated by multiple post-translational modifications, including ubiquitination, yet the molecular mechanism underpinning its reversible deubiquitination remains poorly defined. Here, we identified ubiquitin-specific protease 19 (USP19) as a positive regulator of autophagy, but a negative regulator of type I interferon (IFN) signaling.USP19 stabilizes Beclin-1 by removing the K11-linked ubiquitin chains of Beclin-1 at lysine 437. Moreover, we foundthat USP19 negatively regulates type IIFNsignaling pathway, by blockingRIG-I-MAVSinteraction in a Beclin-1-dependent manner. Depletion of eitherUSP19 or Beclin-1 inhibits autophagic flux and promotes type IIFNsignaling as well as cellular antiviral immunity. Our findings reveal novel dual functions of theUSP19-Beclin-1 axis by balancing autophagy and the production of type IIFNs.

NLRX1 negatively regulates TLR-induced NF-κB signaling by targeting TRAF6 and IKK.

Tight regulation of NF-κB signaling is essential for innate and adaptive immune responses, yet the molecular mechanisms responsible for its negative regulation are not completely understood. Here, we report that NLRX1, a NOD-like receptor family member, negatively regulates Toll-like receptor-mediated NF-κB activation. NLRX1 interacts with TRAF6 or IκB kinase (IKK) in an activation signal-dependent fashion. Upon LPS stimulation, NLRX1 is rapidly ubiquitinated, disassociates from TRAF6, and then binds to the IKK complex, resulting in inhibition of IKKα and IKKß phosphorylation and NF-κB activation. Knockdown of NLRX1 in various cell types markedly enhances IKK phosphorylation and the production of NF-κB-responsive cytokines after LPS stimulation. We further provide in vivo evidence that NLRX1 knockdown in mice markedly enhances susceptibility to LPS-induced septic shock and plasma IL-6 level. Our study identifies a previously unrecognized role for NLRX1 in the negative regulation of TLR-induced NF-κB activation by dynamically interacting with TRAF6 and the IKK complex.

Comparison of Mismatch Repair Status Between Primary and Matched Metastatic Sites in Patients With Colorectal Cancer.

BACKGROUND: Differences between the features of primary cancer and matched metastatic cancer have recently drawn attention in research. This study investigated the concordance in microsatellite instability (MSI) and mismatch repair (MMR) status between primary and corresponding metastatic colorectal cancer (CRC). METHODS: Consecutive patients with metastatic CRC who had both primary and metastatic tumors diagnosed at our institution in January 2008 through December 2016 were identified. Immunohistochemistry was used to test the MMR status of both primary and matched metastatic tumors, and PCR analysis was performed to test MSI in patients with deficient MMR (dMMR) status. RESULTS: A total of 369 patients were included. Of the 46 patients with MSI-high primary tumors, 37 (80.4%) also had MSI-high metastatic tumors, whereas 9 (19.6%) had microsatellite stable (MSS) metastatic tumors. A high concordance was found in patients with liver, lung, or distant lymph node metastases. Interestingly, the discrepancy was more likely to be limited to peritoneal (5/20) or ovarian (4/4) metastasis (chi-square test, P<.001). These organ-specific features were also found in the pooled analysis. Along with the change of MSI-high in primary cancer to MSS in metastatic cancer, lymphocyte infiltration decreased significantly (P=.008). However, the change did not influence survival; the median overall survival of MSI-high and MSS metastatic tumors was 21.3 and 21.6 months, respectively (P=.774). The discrepancy rate was 1.6% for patients with proficient MMR primary tumors. CONCLUSIONS: For patients with dMMR primary tumors, the concordance of MSI and MMR status in primary CRC and corresponding metastatic cancer is potentially organ-specific. High concordance is found in liver, lung, and distant lymph node metastases, whereas discrepancy is more likely to occur in peritoneal or ovarian metastasis. Rebiopsy to evaluate MSI-high/dMMR status might be needed during the course of anti-PD-1 therapy in cases of peritoneal or ovarian metastasis.

Autophagy Regulation of Mammalian Immune Cells.

Autophagy is a fully competent cellular machinery able to carry out the clearance of macromolecules via fusion with the lysosome. Many studies conducted in recent years have revealed that autophagy not only plays a critical role in maintaining cell homeostasis, but can also promote bacterial elimination. Additionally, autophagy exists in most eukaryotic cells including immune cells, such as lymphocytes, neutrophils, eosinophils, mast cells, and natural killer cells. Presently, there are numerous studies focusing on the roles of autophagy in regulating immune response. Autophagy regulates the innate and adaptive immunity by modulating cell differentiation, survival, phagocytosis, antigen presentation, degranulation, and cytokine production. In this chapter, we will summarize how autophagy participates explicitly in the survival and function of the mammalian adaptive and innate immune cells.

Comparison of immunological characteristics between paired mismatch repair-proficient and -deficient colorectal cancer patients.

BACKGROUND: Currently, mismatch repair-deficient (dMMR) status is a promising candidate for targeted immune checkpoint inhibition therapy in colorectal cancer (CRC) patients, however, the potential immunological mechanism has not yet been well clarified and some other predictors need to be excavated as well. METHODS: We collected 330 CRC patients by the match of mismatch repair-proficient (167) and dMMR (163), explored the relationship between MMR status and some important immune molecules including MHC class I, CD3, CD4, CD8, CD56, programmed death-1 and programmed death ligand-1, and investigated the risk factors for dMMR status as well as low MHC class I expression. The Pearson Chi square test was used for analyzing the associations between clinicopathological and immune characteristics and MMR status, and two categories logistic regression model was used for univariate and multivariate analysis to predict the odds ratio of risk factors for dMMR status and low MHC class I expression. RESULTS: Multivariate logistic regression analysis showed that low MHC class I and CD4 expression and high CD8 expression were significant risk factors for dMMR status [odds ratio (OR) = 24.66, 2.94 and 2.97, respectively; all p < 0.05] and dMMR status was the only risk factor for low MHC class I expression (OR = 15.34; p < 0.001). CONCLUSIONS: High CD8 and low MHC class I expression suggests the contradiction and complexity of immune microenvironment in dMMR CRC patients. Some other immunocytes such as CD56+ cells might also participate in the process of immune checkpoint inhibition, whereas needs further investigations.

Second primary colorectal cancer after the initial primary colorectal cancer.

BACKGROUND: Initial primary colorectal cancer (IPCRC) has a high risk of developing into second primary colorectal cancer (SPCRC). Right-sided colon cancer (RCC) and left-sided colon cancer (LCC) have different characteristics and are considered to be two different entities. However, the different risks for SPCRC in categorized tumor sites and SPCRC subcategorized sites have not been fully elucidated to date. We aimed to compare incidence and survival of IPCRC and SPCRC and characterize the risk factors of SPCRC while also comparing the different SPCRC characteristics. METHODS: We used the National Cancer Institute's Surveillance, Epidemiology, and End Results (SEER) data to compute standardized incidence ratios (SIR) in order to estimate risk of SPCRC after IPCRC diagnosis. The most prominent risk factors for SPCRC were measured by multivariate regression analysis and the temporal trend of SPCRC incidence was assessed with Joinpoint regression. Survival of patients with SPCRC and IPCRC was compared by Kaplan-Meier analysis. RESULTS: Patients with IPCRC were 1.73 times more likely to develop SPCRC (SIR = 1.73, 95% CI 1.69-1.78). SPCRC incidence declined since the first 8 years of IPCRC diagnosis to baseline. We demonstrated poorer survival with SPCRC compared with IPCRC while second RCC resulted in better survival compared with second LCC. Black ethnicity, age range 70-79, and LCC were associated with the highest risk of developing SPCRC. CONCLUSION: The characteristic differences between second LCC and RCC were relatively narrow. Furthermore, in those with SPCRC, RCC had the best survival outcome.

Self-assembled nano-balls released from multistage vector for cancer therapy.

The efficacy of cancer drugs is often compromised due to the existence of biological barriers such as nonspecific distribution, hemorheological flow limitation and endothelial extravasation, impaired delivery across tumor cell membranes and tissue, and multidrug resistance. To overcome these obstacles, Xu et al developed an injectable nanoparticle generator platform to negotiate with the biological barriers and enable self-assembly of nano-balls in situ in order to maximize drug accumulation inside the tumor tissues and hence the therapeutic efficacy. This perspective aims to elaborate the designing strategy, and discuss the mechanism of action of the new drug and the potential for future development of nanoparticulate drugs.

Cortisol is associated with low frequency of interleukin 10-producing B cells in patients with atherosclerosis.

It is accepted that inflammation plays a critical role in the development of atherosclerosis; the pathogenesis is not clear. B-cell-produced interleukin (IL) 10 is an immune regulatory cytokine that can inhibit immune inflammation. This study tests a hypothesis that a psychological stress hormone, cortisol, suppresses IL-10 expression in peripheral B cells of patients with atherosclerosis. Peripheral blood samples were collected from patients with coronary artery atherosclerosis. B cells were isolated from the blood samples to be analyzed for the expression of IL-10 and micro RNA (miR) 98 by real-time polymerase chain reaction. We observed that the frequency of IL-10+ B cell was less in patients with atherosclerosis than healthy controls. The serum cortisol levels were higher in the patients than that in healthy controls. Peripheral B-cell frequency was negatively correlated with the serum cortisol levels. Exposure of B cells to cortisol increased the expression of miR-98 in B cells. Cortisol also inhibited the expression of IL-10 in B cells, in which miR-98 played a critical role. Treating B cells from atherosclerosis patients with anti-miR-98 liposomes reversed the ability of expression of IL-10 in the cells. The expression of IL-10 is suppressed in peripheral B cells, which can be up regulated by anti-miR-98 liposomes.

Cancer incidence and mortality in Gansu province, 2012.

OBJECTIVE: Population-based cancer registration data in 2012 from all available cancer registries in Gansu province were collected by the Central Cancer Registry of Gansu. The numbers of new cancer cases and cancer deaths in Gansu province with compiled cancer incidence and mortality rates were estimated. METHODS: In 2015, data from 7 registries in Gansu province were qualified. The pooled data were stratified by area (urban/rural), gender, age group (0, 1.4, 5.9, 10.14, …, 85+) and cancer type. New cancer cases and deaths were estimated using age-specific rates and corresponding population of Gansu province in 2012. The Chinese census data in 2000 and Segi's population were applied for age-standardized rates. All the rates were expressed per 100,000 person-years. RESULTS: Qualified 7 cancer registries (3 urban and 4 rural registries) covered 2,956,560 populations of Gansu province in 2012. The percentage of cases morphologically verified (MV%) and death certificate-only cases (DCO%) were 72.41% and 1.65%, respectively, and the mortality to incidence rate ratio (M/I) was 0.63. It was estimated that there were 575,600 new cancer cases and 331,300 cancer deaths in Gansu province in 2012. The incidence rate was 223.29/100,000 (244.14/100,000 in males and 201.50/100,000 in females), the age-standardized incidence rates by Chinese standard population (ASIRC) and by world standard population (ASIRW) were 208.95/100,000 and 206.41/100,000 with the cumulative incidence rate (0.74 years old) of 22.49%. The crude incidence rate in urban areas was equal to that in rural areas. However, after adjusted by age, the cancer incidence rate in urban was the same as that of rural areas. The crude mortality in Gansu province was 128.54/100,000 (135.04/100,000 in males and 124.43/100,000 in females), the age-standardized mortality rates by Chinese standard population (ASMRC) and by world standard population (ASMRW) were 109.54/100,000 and 108.44/100,000, respectively, and the cumulative mortality rate (0.74 years old) was 12.91%. The crude cancer mortality rate in urban areas was higher than that of rural areas. However, the agestandardized rate in urban areas was the same as that of rural areas. Cancers of stomach, liver, lung, esophagus, female breast, colorectum, cervix, brain, pancreas and ovary were the most common cancers, accounting for about 83.56% of all cancer new cases. Stomach cancer, liver cancer, lung cancer, esophageal cancer, female breast cancer, colorectal cancer, brain cancer, lymphoma, pancreatic cancer and cervix cancer were the leading causes of cancer death, accounting for about 85.58% of all cancer deaths. The burden between urban and rural, males and females was different. CONCLUSIONS: Registration data of Gansu province were qualified to provide basic information on population-based cancer incidence, mortality for cancer prevention and control. The digestive tract cancer burden in Gansu province, especially for males in rural areas, was higher. The incidence rate of female breast cancer was higher in urban areas. Targeted prevention, early detection and treatment programs should be carried out by health department to control the cancer burden.

Recent progress in mRNA cancer vaccines.

The research and development of messenger RNA (mRNA) cancer vaccines have gradually overcome numerous challenges through the application of personalized cancer antigens, structural optimization of mRNA, and the development of alternative RNA-based vectors and efficient targeted delivery vectors. Clinical trials are currently underway for various cancer vaccines that encode tumor-associated antigens (TAAs), tumor-specific antigens (TSAs), or immunomodulators. In this paper, we summarize the optimization of mRNA and the emergence of RNA-based expression vectors in cancer vaccines. We begin by reviewing the advancement and utilization of state-of-the-art targeted lipid nanoparticles (LNPs), followed by presenting the primary classifications and clinical applications of mRNA cancer vaccines. Collectively, mRNA vaccines are emerging as a central focus in cancer immunotherapy, offering the potential to address multiple challenges in cancer treatment, either as standalone therapies or in combination with current cancer treatments.