Role of miR-106-mediated mitogen-activated protein kinase signaling pathway in oxidative stress injury and inflammatory infiltration in the liver of the mouse with gestational hypertension.

We aim to investigate the role of miR-106-mediated mitogen-activated protein kinase (MAPK) signaling pathway in oxidative stress (OS) injury and inflammatory infiltration in the liver of the mouse with gestational hypertension (GH). Ninety specific pathogen-free mice (Kunming species) during middle to late gestation were selected for the study. Fifteen mice were used as control, while the rest were used for establishing the GH model. The mice were assigned to six groups: normal group (normal gestation), model group (GH model), negative control group (GH model, intravenously injected with negative control vector), miR-106a-mimic group (GH model, intravenously injected with vector overexpressing miR-106a, which mimics the overexpression of endogenous mature miR-106a), SB203580 group (GH model, intravenously injected with MAPK pathway inhibitor SB203580), and miR-106a-mimic+SB203580 group (GH model, intravenously injected with SB203580 and vector overexpressing miR-106a). Fourteen days after electrical stimulation, all the groups except for the normal group had elevated blood pressure vs those on day 0 and 7. Compared with the normal group, the other groups had lower levels of miR-106a expression, nitric oxide, nitric oxide synthase, catalase, superoxide dismutase, S cell ratio, and interleukin-4 (IL-4) and IL-10 in the serum and liver as opposed to increased levels of blood pressure, p38MAPK mRNA expression, p-p38MAPK positive expression rate, protein expressions of p-p38MAPK, p-ERK, and p-JNK, H2 O2 and malondialdehyde in liver, G0/G1 cell ratio, apoptosis rate, and IL-6, interferon-γ (IFN-γ), and IFN-α in the serum and liver (all P < .05). The miR-106 overexpression or inhibiting MAPK signaling pathway can attenuate OS injury and inflammatory response in the liver of the mouse with GH, and the effect can be even better if both miR-106a overexpression and inhibiting MAPK pathway are applied. In conclusion, miR-106a overexpression can inhibit OS injury and inflammatory infiltration in the liver of the mouse with GH by mediating MAPK signaling pathway.

NLRP6 deficiency suppresses colorectal cancer liver metastasis growth by modulating M-MDSC-induced immunosuppressive microenvironment.

Colorectal cancer liver metastasis (CRLM) a profound influence on the prognosis of patients with colorectal cancer (CRC), prompting a comprehensive inquiry into its underlying mechanisms. Amidst the multifaceted tumor microenvironment, myeloid-derived suppressor cells (MDSCs) have emerged as pivotal orchestrators of immune modulation. However, their specific contributions to the CRLM have not been explored. The role of NLRP6, a member of the NOD-like receptor family, is of interest. Employing a liver metastasis model, our investigation revealed a heightened accumulation of monocytic MDSCs (M-MDSCs) within metastatic sites, culminating in an immunosuppressive milieu characterized by depleted CD8+ T cell populations. Remarkably, the absence of NLRP6 disrupts this intricate immunosuppressive network, highlighting its nuanced role in sculpting the trajectory of CRLM. This study elucidates the interplay between NLRP6 and MDSCs, potentially guiding novel therapeutic strategies to recalibrate the immune microenvironment in CRLM and enhance patient outcomes.

A combination influenza mRNA vaccine candidate provided broad protection against diverse influenza virus challenge.

Influenza viruses present a significant threat to global health. The production of a universal vaccine is considered essential due to the ineffectiveness of current seasonal influenza vaccines against mutant strains. mRNA technology offers new prospects in vaccinology, with various candidates for different infectious diseases currently in development and testing phases. In this study, we encapsulated a universal influenza mRNA vaccine. The vaccine encoded influenza hemagglutinin (HA), nucleoprotein (NP), and three tandem repeats of matrix protein 2 (3M2e). Twice-vaccinated mice exhibited strong humoral and cell-mediated immune responses in vivo. Notably, these immune responses led to a significant reduction in viral load of the lungs in challenged mice, and also conferred protection against future wild-type H1N1, H3N2, or H5N1 influenza virus challenges. Our findings suggest that this mRNA-universal vaccine strategy for influenza virus may be instrumental in mitigating the impact of future influenza pandemics.

An Engineered Influenza a Virus Expressing the Co-Stimulator OX40L as an Oncolytic Agent Against Hepatocellular Carcinoma.

Background: Oncolytic virus (OV) therapy has emerged as a promising novel form of immunotherapy. Moreover, an increasing number of studies have shown that the therapeutic efficacy of OV can be further improved by arming OVs with immune-stimulating molecules. Methods: In this study, we used reverse genetics to produce a novel influenza A virus, termed IAV-OX40L, which contained the immune-stimulating molecule OX40L gene in the influenza virus nonstructural (NS1) protein gene. The oncolytic effect of IAV-OX40L was explored on hepatocellular carcinoma (HCC)HCC cells in vitro and in vivo. Results: Hemagglutination titers of the IAV-OX40L virus were stably 27-28 in specific-pathogen-free chicken embryos. The morphology and size distribution of IAV-OX40L are similar to those of the wild-type influenza. Expression of OX40L protein was confirmed by Western blot and immunofluorescence. MTS assays showed that the cytotoxicity of IAV-OX40L was higher in HCC cells (HepG2 and Huh7) than in normal liver cells (MIHA) in a time- and dose-dependent manner in vitro. We found that intratumoral injection of IAV-OX40L reduced tumor growth and increased the survival rate of mice compared with PR8-treated controls in vivo. In addition, the pathological results showed that IAV-OX40L selectively destroyed tumor tissues without harming liver and lung tissues. CD4+ and CD8+ T cells of the IAV-OX40L group were significantly increased in the splenic lymphocytes of mice. Further validation confirmed that IAV-OX40L enhanced the immune response mainly by activating Th1-dominant immune cells, releasing interferon-γ and interleukin-2. Conclusion: Taken together, our findings demonstrate the novel chimeric influenza OV could provide a potential therapeutic strategy for combating HCC and improve the effectiveness of virotherapy for cancer therapy.

A Recombinant Oncolytic Influenza Virus Carrying GV1001 Triggers an Antitumor Immune Response.

Oncolytic viruses are able to lyse tumor cells selectively in the liver without killing normal hepatocytes, in addition to activating the immune response. Oncolytic virus therapy is expected to revolutionize the treatment of liver cancer, including hepatocellular carcinoma (HCC), one of the most frequent and fatal malignancies. In this study, reverse genetics techniques were exploited to load NA fragments of the A/PuertoRico/8/34 virus (PR8) with GV1001 peptides derived from human telomerase reverse transcriptase. An in vitro assessment of the therapeutic effect of the recombinant oncolytic virus was followed by an in vivo study in mice with HCC. The recombinant virus was verified by sequencing of the recombinant viral gene sequence, and viral virulence was detected by hemagglutination assays and based on the 50% tissue culture infectious dose (TCID50). The morphological structure of the virus was observed by electron microscopy, and GV1001 peptide was localized by cellular immunofluorescence. The selective cytotoxicity of the recombinant oncolytic virus in vitro was demonstrated in cultured HCC cells and normal hepatocytes, as only the tumor cells were killed; the normal cells were not significantly altered. Consistent with the in vitro results, the recombinant oncolytic influenza virus significantly inhibited liver tumor growth in mice in vivo, in addition to inducing an antitumor immune response, including an increase in the number of CD4+ and CD8+ T lymphocytes and, in turn, improving survival. Our results suggest that oncolytic influenza virus carrying GV1001 is a promising immunotherapy in patients with HCC.

Enhanced NK cell activation via eEF2K-mediated potentiation of the cGAS-STING pathway in hepatocellular carcinoma.

BACKGROUND: Liver cancer, particularly hepatocellular carcinoma (HCC), is characterized by a high mortality rate, attributed primarily to the establishment of an immunosuppressive microenvironment. Within this context, we aimed to elucidate the pivotal role of eukaryotic elongation factor 2 kinase (eEF2K) in orchestrating the infiltration and activation of natural killer (NK) cells within the HCC tumor microenvironment. By shedding light on the immunomodulatory mechanisms at play, our findings should clarify HCC pathogenesis and help identify potential therapeutic intervention venues. METHODS: We performed a comprehensive bioinformatics analysis to determine the functions of eEF2K in the context of HCC. We initially used paired tumor and adjacent normal tissue samples from patients with HCC to measure eEF2K expression and its correlation with prognosis. Subsequently, we enrolled a cohort of patients with HCC undergoing immunotherapy to examine the ability of eEF2K to predict treatment efficacy. To delve deeper into the mechanistic aspects, we established an eEF2K-knockout cell line using CRISPR/Cas9 gene editing. This step was crucial for verifying activation of the cGAS-STING pathway and the subsequent secretion of cytokines. To further elucidate the role of eEF2K in NK cell function, we applied siRNA-based techniques to effectively suppress eEF2K expression in vitro. For in vivo validation, we developed a tumor-bearing mouse model that enabled us to compare the infiltration and activation of NK cells within the tumor microenvironment following various treatment strategies. RESULTS: We detected elevated eEF2K expression within HCC tissues, and this was correlated with an unfavorable prognosis (30.84 vs. 20.99 months, P = 0.033). In addition, co-culturing eEF2K-knockout HepG2 cells with dendritic cells led to activation of the cGAS-STING pathway and a subsequent increase in the secretion of IL-2 and CXCL9. Moreover, inhibiting eEF2K resulted in notable NK cell proliferation along with apoptosis reduction. Remarkably, after combining NH125 and PD-1 treatments, we found a significant increase in NK cell infiltration within HCC tumors in our murine model. Our flow cytometry analysis revealed reduced NKG2A expression and elevated NKG2D expression and secretion of granzyme B, TNF-α, and IFN-γ in NK cells. Immunohistochemical examination confirmed no evidence of damage to vital organs in the mice treated with the combination therapy. Additionally, we noted higher levels of glutathione peroxidase and lipid peroxidation in the peripheral blood serum of the treated mice. CONCLUSION: Targeted eEF2K blockade may result in cGAS-STING pathway activation, leading to enhanced infiltration and activity of NK cells within HCC tumors. The synergistic effect achieved by combining an eEF2K inhibitor with PD-1 antibody therapy represents a novel and promising approach for the treatment of HCC.

Three-dimensional chromatin landscapes in hepatocellular carcinoma associated with hepatitis B virus.

BACKGROUND: Three-dimensional (3D) chromatin architecture frequently altered in cancer. However, its changes during the pathogenesis of hepatocellular carcinoma (HCC) remained elusive. METHODS: Hi-C and RNA-seq were applied to study the 3D chromatin landscapes and gene expression of HCC and ANHT. Hi-C Pro was used to generate genome-wide raw interaction matrices, which were normalized via iterative correction (ICE). Moreover, the chromosomes were divided into different compartments according to the first principal component (E1). Furthermore, topologically associated domains (TADs) were visualized via WashU Epigenome Browser. Furthermore, differential expression analysis of ANHT and HCC was performed using the DESeq2 R package. Additionally, dysregulated genes associated with 3D genome architecture altered were confirmed using TCGA, qRT-PCR, immunohistochemistry (IHC), etc. RESULTS: First, the intrachromosomal interactions of chr1, chr2, chr5, and chr11 were significantly different, and the interchromosomal interactions of chr4-chr10, chr13-chr21, chr15-chr22, and chr16-chr19 are remarkably different between ANHT and HCC, which resulted in the up-regulation of TP53I3 and ZNF738 and the down-regulation of APOC3 and APOA5 in HCC. Second, 49 compartment regions on 18 chromosomes have significantly switched (A-B or B-A) during HCC tumorigenesis, contributing to up-regulation of RAP2A. Finally, a tumor-specific TAD boundary located on chr5: 6271000-6478000 and enhancer hijacking were identified in HCC tissues, potentially associated with the elevated expression of MED10, whose expression were associated with poor prognosis of HCC patients. CONCLUSION: This study demonstrates the crucial role of chromosomal structure variation in HCC oncogenesis and potential novel biomarkers of HCC, laying a foundation for cancer precision medicine development.

Pilot study of blood perfusion coherence along the meridian in forearm.

BACKGROUND: Many studies have explored the relationship between skin microcirculation and meridian activation. However, few studies have examined blood perfusion coherence along the meridians, and other studies have suggested that the skin vasodilator response relates to age. This study investigated blood perfusion coherence characteristics along the meridian of the forearm in healthy volunteers. METHODS: A total of 15 young subjects (25.53 ± 2.20) and 15 middle-aged subjects (50.07 ± 3.37) were recruited for this study. Before experiments, each subject was placed in a temperature-controlled room for 60 min. Skin blood perfusion from five points was recorded simultaneously using a full-field laser perfusion imager before and after inflatable occlusion. The five points comprised three points located on the pericardium meridian, and two points from different locations. Coherence analysis between these points was performed at different frequency intervals from 0.0095 to 2 Hz. RESULTS: In young subjects, the coherence value was unchanged before and after occlusion, and there was no significant difference in coherence value between meridian-meridian points (M-M) and meridian-parameridian points (M-P). In middle-aged subjects, the coherence value increased significantly in both M-M and M-P at frequency intervals of 0.14-0.4 Hz, 0.4-1.6 Hz, and 1.6-2 Hz. However, there was no significant difference in coherence values between M-M and M-P. CONCLUSIONS: Inflatable occlusion can increase middle-aged subjects' blood perfusion coherence value of the forearm. However, there is no specificity in meridian location.

Spotlight on NLRP6 and Tumor Research Situation: A Potential Cancer Participant.

NOD-like receptor family pyrin domain containing 6 (NLRP6) is a new pattern recognition receptor in the mammalian innate immune system. Both the liver and the gut exhibit substantial levels of cytoplasmic expression. It can speed up cell response to endogenous danger signals or exogenous pathogen infection. NLRP6 can function in various ways as an inflammasome or a noninflammasome. The understanding of NLRP6 is steadily increasing thanks to ongoing investigations, but due to discrepancies in how those studies have described their link with tumors, the significance of NLRP6 in the emergence of cancer is still debatable as of this writing. This article will use the structure and function of NLRP6 as the pivotal point and thoroughly explain the present interactions between NLRP6 and tumors and any possible clinical benefits.

Mechanism of Bazhen decoction in the treatment of colorectal cancer based on network pharmacology, molecular docking, and experimental validation.

Objective: Bazhen Decoction (BZD) is a common adjuvant therapy drug for colorectal cancer (CRC), although its anti-tumor mechanism is unknown. This study aims to explore the core components, key targets, and potential mechanisms of BZD treatment for CRC. Methods: The Traditional Chinese Medicine Systems Pharmacology (TCMSP) was employed to acquire the BZD's active ingredient and targets. Meanwhile, the Drugbank, Therapeutic Target Database (TTD), DisGeNET, and GeneCards databases were used to retrieve pertinent targets for CRC. The Venn plot was used to obtain intersection targets. Cytoscape software was used to construct an "herb-ingredient-target" network and identify core targets. GO and KEGG pathway enrichment analyses were conducted using R language software. Molecular docking of key ingredients and core targets of drugs was accomplished using PyMol and Autodock Vina software. Cell and animal research confirmed Bazhen Decoction efficacy and mechanism in treating colorectal cancer. Results: BZD comprises 173 effective active ingredients. Using four databases, 761 targets related to CRC were identified. The intersection of BZD and CRC yielded 98 targets, which were utilized to construct the "herb-ingredient-target" network. The four key effector components with the most targets were quercetin, kaempferol, licochalcone A, and naringenin. Protein-protein interaction (PPI) analysis revealed that the core targets of BZD in treating CRC were AKT1, MYC, CASP3, ESR1, EGFR, HIF-1A, VEGFR, JUN, INS, and STAT3. The findings from molecular docking suggest that the core ingredient exhibits favorable binding potential with the core target. Furthermore, the GO and KEGG enrichment analysis demonstrates that BZD can modulate multiple signaling pathways related to CRC, like the T cell receptor, PI3K-Akt, apoptosis, P53, and VEGF signaling pathway. In vitro, studies have shown that BZD dose-dependently inhibits colon cancer cell growth and invasion and promotes apoptosis. Animal experiments have shown that BZD treatment can reverse abnormal expression of PI3K, AKT, MYC, EGFR, HIF-1A, VEGFR, JUN, STAT3, CASP3, and TP53 genes. BZD also increases the ratio of CD4+ T cells to CD8+ T cells in the spleen and tumor tissues, boosting IFN-γ expression, essential for anti-tumor immunity. Furthermore, BZD has the potential to downregulate the PD-1 expression on T cell surfaces, indicating its ability to effectively restore T cell function by inhibiting immune checkpoints. The results of HE staining suggest that BZD exhibits favorable safety profiles. Conclusion: BZD treats CRC through multiple components, targets, and metabolic pathways. BZD can reverse the abnormal expression of genes such as PI3K, AKT, MYC, EGFR, HIF-1A, VEGFR, JUN, STAT3, CASP3, and TP53, and suppresses the progression of colorectal cancer by regulating signaling pathways such as PI3K-AKT, P53, and VEGF. Furthermore, BZD can increase the number of T cells and promote T cell activation in tumor-bearing mice, enhancing the immune function against colorectal cancer. Among them, quercetin, kaempferol, licochalcone A, naringenin, and formaronetin are more highly predictive components related to the T cell activation in colorectal cancer mice. This study is of great significance for the development of novel anti-cancer drugs. It highlights the importance of network pharmacology-based approaches in studying complex traditional Chinese medicine formulations.

Evaluate laser needle effect on blood perfusion signals of contralateral hegu acupoint with wavelet analysis.

Our previous studies suggested that the MBF in contralateral Hegu acupoint (IL4) increased after ipsilateral Hegu acupoint was stimulated with manual acupuncture. In this study, twenty-eight (28) healthy volunteers were recruited and were randomly divided into Hegu acupoint stimulation group and Non-Hegu stimulation group. All subjects received the same model stimulation of the laser needle for 30 min in right Hegu acupoint and Non-Hegu acupoint, respectively. MBF of left LI4 was measured by the laser Doppler perfusion imaging system. The original data dealt with morlet wavelet analysis and the average amplitude and power spectral density of different frequency intervals was acquired. The results indicated that right Hegu stimulation with the laser needle might result in the increase of left Hegu acupoint MBF. 40 min later after ceased stimulation, the MBF is still increasing significantly, whereas the MBF has no significantly change in Non-Hegu stimulation group. The wavelet analysis result suggested that compared to Non-Hegu stimulation, stimulated to right Hegu acupoint might result in the increase of average amplitude in frequency intervals of 0.0095-0.02 Hz, 0.02-0.06 Hz, and 0.06-0.15 Hz, which might be influenced by the endothelial, neurogenic, and the intrinsic myogenic activity of the vessel wall, respectively.

A transcontinental pilot study for acupuncture lifting-thrusting and twisting-rotating manipulations.

The goal of this study was to observe possible changes of the skin microvascular perfusion on the acupoints and related areas and to quantify influences of acupuncture stimulation on the volunteers' blood pressure, heart rate, and heart rate variability (HRV). During the measurement, the needling sensations of volunteers were enquired and recorded. Ten healthy volunteers with a mean age ± SD of 25.4 ± 2.6 years were enrolled, and acupuncture stimulation was performed on ST36 (Zusanli, right side), in pure lifting-thrusting or twisting-rotating manipulation. During needling, we observed the changing of microvascular perfusion on ST36, 37, 38, and a control point using MOOR speckle laser blood flow scanning. Electrocardiogram and blood pressure were registered before, during, and after needling. Both lifting-thrusting and twisting-rotating needle manipulations could decrease blood pressure and heart rate while improving HRV significantly. There were significant differences in microvascular perfusion on acupoints ST36, 37, 38, and the control point following these two kinds of needle manipulation. The needling sensation caused by lifting-thrusting is stronger than that of twisting-rotating manipulation. Significant differences between lifting-thrusting and twisting-rotating acupuncture stimulation methods show that the mechanisms may be different and need to be researched thoroughly in the future.

Observation of Microvascular Perfusion in the Hegu (LI4) Acupoint Area after Deqi Acupuncture at Quchi (LI11) Acupoint Using Speckle Laser Blood Flow Scanning Technology.

The aim of this study was to investigate the traditional meridian theory using speckle laser blood flow scanning technology to observe microcirculation of the Hegu acupoint area after acupuncture stimulation on distant points. An observational study was conducted to observe the microvascular perfusion of Hegu (LI4) and control points after acupuncturing Quchi (LI11). Thirty healthy volunteers (mean age 31.6 ± 8.7 years) received deqi acupuncture on Quchi (LI11, right side), and simultaneously changes in microvascular perfusion of Sanjian (LI3), Hegu (LI4), Yangxi (LI5), and two control points were observed before, during, and after needling using a MOOR speckle laser. The results showed that the changes in microvascular perfusion of the observed points are not regular. After correction, the experiment showed that the blood perfusion on 3 meridian acupoints was increased while the perfusion on 2 control points was decreased following acupuncture stimulation, the changes at Hegu (LI4) being the statistically most significant ones. Deqi acupuncture can help in regulating the body's blood flow, with a certain degree of meridian specificity.

The influence of different methods of acupuncture on skin surface perfusion.

OBJECTIVE: To compare the effects on the body surface of different kinds of placebo/sham acupuncture with that of traditional needling sensation acupuncture. METHODS: Point Neiguan (PC 6) of 29 healthy subjects, 19 males and 10 females, was stimulated with needling sensation, shallow, placebo and deep acupuncture at the non-acupoint. After stimulation with different methods of acupuncture, the change in perfusion of the micrangium in the skin surface around the elbow joint, with Point Quze (PC 3) as its center, was observed with laser Doppler blood-flow imaging. RESULTS: Judging from the absolute value of perfusion of the skin surface micrangium, several methods of acupuncture can cause change in blood flow. The ratio of blood-flow perfusion in the meridian area in and around Quze declined before and after needling insertion in needling sensation acupuncture and shallow acupuncture. This observation did not occur in placebo and non-acupoint acupuncture. Needling sensation acupuncture at an acupoint can relatively reduce the perfusion of the micrangium in the projective area of the meridian where the acupoint is located on the body surface (P < 0.05), indicating the specificity of meridians. CONCLUSIONS: Stimulation of an acupoint or a point on the body surface with any type of acupuncture can cause change in blood flow in the skin near the needling point. However, the biological mechanism underlying this phenomenon needs to be further explored.

mRNA vaccines: A novel weapon to control infectious diseases.

Infectious diseases have always threatened human life, but with the development of vaccines, effective strategies for preventing and controlling these diseases have become available. The global outbreak of COVID-19 ushered in the advent of mRNA vaccine technologies, which quickly led to the introduction of mRNA vaccines effective against SARS-CoV-2. The success of this approach has stimulated research into the use of mRNA vaccines in the fight against other emerging as well as remerging infectious diseases. This review examines the constructive strategies and delivery systems used in mRNA vaccines and provides an overview of current clinical trials of those vaccines in the prevention of infectious diseases. The underlying mechanisms of mRNA vaccines are also discussed, including the double-edged sword of the innate immune response. Finally, the challenges but also the potential of mRNA vaccines are considered.

mRNA Vaccines: The Dawn of a New Era of Cancer Immunotherapy.

mRNA therapy is a novel anticancer strategy based on in vitro transcription (IVT), which has potential for the treatment of malignant tumors. The outbreak of the COVID-19 pandemic in the early 21st century has promoted the application of mRNA technologies in SARS-CoV-2 vaccines, and there has been a great deal of interest in the research and development of mRNA cancer vaccines. There has been progress in a number of key technologies, including mRNA production strategies, delivery systems, antitumor immune strategies, etc. These technologies have accelerated the progress and clinical applications of mRNA therapy, overcoming problems encountered in the past, such as instability, inefficient delivery, and weak immunogenicity of mRNA vaccines. This review provides a detailed overview of the production, delivery systems, immunological mechanisms, and antitumor immune response strategies for mRNA cancer vaccines. We list some mRNA cancer vaccines that are candidates for cancer treatment and discuss clinical trials in the field of tumor immunotherapy. In addition, we discuss the immunological mechanism of action by which mRNA vaccines destroy tumors as well as challenges and prospects for the future.

Data-independent acquisition mass spectrometry identification of extracellular vesicle biomarkers for gastric adenocarcinoma.

Early diagnosis of gastric adenocarcinoma (GAC) can effectively prevent the progression of the disease and significantly improve patient survival. Currently, protein markers in clinical practice barely meet patient needs; it is therefore imperative to develop new diagnostic biomarkers with high sensitivity and specificity. In this study, we extracted extracellular vesicles (EV) from the sera of 33 patients with GAC and 19 healthy controls, then applied data-independent acquisition (DIA) mass spectrometry to measure protein expression profiles. Differential protein expression analysis identified 23 proteins showing expression patterns across different cancer stages, from which 15 proteins were selected as candidate biomarkers for GAC diagnosis. From this subset of 15 proteins, up to 6 proteins were iteratively selected as features and logistic regression was used to distinguish patients from healthy controls. Furthermore, serum-derived EV from a new cohort of 12 patients with gastric cancer and 18 healthy controls were quantified using the same method. A classification panel consisting of GSN, HP, ORM1, PIGR, and TFRC showed the best performance, with a sensitivity and negative predictive value (NPV) of 0.83 and 0.82. The area under curve (AUC) of the receiver operating characteristic (ROC) is 0.80. Finally, to facilitate the diagnosis of advanced stage GAC, we identified a 3-protein panel consisting of LYZ, SAA1, and F12 that showed reasonably good performance with an AUC of 0.83 in the validation dataset. In conclusion, we identified new protein biomarker panels from serum EVs for early diagnosis of gastric cancer that worth further validation.

An OX40L mRNA vaccine inhibits the growth of hepatocellular carcinoma.

mRNA cancer vaccines show therapeutic potential for malignant tumors, including hepatocellular carcinoma (HCC). We optimized and synthesized stable mRNA encoding costimulator Oxford 40 ligand (OX40L). For systemic delivery, OX40L mRNAs were loaded into lipid nanoparticles (LNPs). The expression and costimulatory effects of OX40L were investigated in vitro. OX40L was expressed on the cell surface and costimulated T cells. In vivo, intratumoral injection of LNPs encapsulating OX40L mRNAs significantly reduced tumor growth and increased the survival of mice bearing H22 tumors. Importantly, CD4+ and CD8+ T cells were significantly increased in the OX40L mRNA group in vivo. Taken together, our findings provide a promising clinical strategy for immunotherapy for HCC using mRNA vaccines.

Comparison of the effects of traditional box-moxibustion and eletrothermal Bian-stone moxibustion on volume of blood flow in the skin.

OBJECTIVE: To investigate and compare the effects of Electrothermal Bian-stone moxibustion and traditional box-moxibustion on capillary blood perfusion in the body surface. METHODS: Twenty healthy subjects, 18 females and 2 males, aged averagely 36 years, were enrolled in this experiment. The traditional box-moxibustion and electrothermal Bian-stone moxibustion were respectively given on the hypogastrium around Guanyuan (CV 4), and the changes of local capillary blood perfusion in the body surface were investigated with Laser Doppler Perfusion Imaging (LDPI). RESULTS: The traditional box-moxibustion was superior to the electrothermal Bian-stone moxibustion in the transient increasing effect in whole body and the long-term increasing effect in the local area of moxibustion on capillary blood perfusion; but for both methods, the increasing effect on the capillary blood perfusion in the local area of moxibustion was same. CONCLUSION: Electrothermal Bian-stone moxibustion can not completely take the place of the traditional box-moxibustion; but for some disorders with local pain, they possibly have the same effects.

Proteomic analysis of lung cancer cells reveals a critical role of BCAT1 in cancer cell metastasis.

Metastasis is the major cause of high mortality in lung cancer. Exploring the underlying mechanisms of metastasis thus holds promise for identifying new therapeutic strategies that may enhance survival. Methods: We applied quantitative mass spectrometry to compare protein expression profiles between primary and metastatic lung cancer cells whilst investigating metastasis-related molecular features. Results: We discovered that BCAT1, the key enzyme in branched-chain amino acid metabolism, is overexpressed at the protein level in metastatic lung cancer cells, as well as in metastatic tissues from lung cancer patients. Analysis of transcriptomic data available in the TCGA database revealed that increased BCAT1 transcription is associated with poor overall survival of lung cancer patients. In accord with a critical role in metastasis, shRNA-mediated knockdown of BCAT1 expression reduced migration of metastatic cells in vitro and the metastasis of these cells to distal organs in nude mice. Mechanistically, high levels of BCAT1 depleted α-ketoglutarate (α-KG) and promoted expression of SOX2, a transcription factor regulating cancer cell stemness and metastasis. Conclusion: Our findings suggest that BCAT1 plays an important role in promoting lung cancer cell metastasis, and may define a novel pathway to target as an anti-metastatic therapy.