Hepatoprotective effect of diammonium glycyrrhizinate and neuroprotective effect of piperazine ferulate on AmB-induced liver and kidney injury by suppressing apoptosis in vitro and in vivo.

Amphotericin B (AmB) induced liver and kidney injury is often responsible for hepatic and renal dysfunction. Therefore, the protection strategy on liver and renal functions in patients treated with AmB should be emphasized. In this paper, diammonium glycyrrhizinate (DG) and piperazine ferulate (PF) were taken as the research object to study its hepatoprotective and neuroprotective effect on AmB-induced liver and kidney damage in vitro and in vivo. The microplate method and ELISA kits were employed for the biochemical detection in the serum and urine of mice. Flow cytometric analysis and western blotting analysis were conducted to study the mechanism of DG and PF. Our results confirmed the prevention capacity of DG and PF on AmB-induced liver and kidney injury through the alleviation of pathological changes and enzyme reducing action. Furthermore, DG and PF suppressed ROS-mediated mitochondrial apoptosis in AmB-treated mice and cells through Caspase pathway and Caspase-independent AIF pathway. In summary, DG and PF could protect AmB-induced hepatotoxicity and nephrotoxicity by disrupting oxidative stress and apoptosis.

Application of AI in Multilevel Pain Assessment Using Facial Images: Systematic Review and Meta-Analysis.

BACKGROUND: The continuous monitoring and recording of patients' pain status is a major problem in current research on postoperative pain management. In the large number of original or review articles focusing on different approaches for pain assessment, many researchers have investigated how computer vision (CV) can help by capturing facial expressions. However, there is a lack of proper comparison of results between studies to identify current research gaps. OBJECTIVE: The purpose of this systematic review and meta-analysis was to investigate the diagnostic performance of artificial intelligence models for multilevel pain assessment from facial images. METHODS: The PubMed, Embase, IEEE, Web of Science, and Cochrane Library databases were searched for related publications before September 30, 2023. Studies that used facial images alone to estimate multiple pain values were included in the systematic review. A study quality assessment was conducted using the Quality Assessment of Diagnostic Accuracy Studies, 2nd edition tool. The performance of these studies was assessed by metrics including sensitivity, specificity, log diagnostic odds ratio (LDOR), and area under the curve (AUC). The intermodal variability was assessed and presented by forest plots. RESULTS: A total of 45 reports were included in the systematic review. The reported test accuracies ranged from 0.27-0.99, and the other metrics, including the mean standard error (MSE), mean absolute error (MAE), intraclass correlation coefficient (ICC), and Pearson correlation coefficient (PCC), ranged from 0.31-4.61, 0.24-2.8, 0.19-0.83, and 0.48-0.92, respectively. In total, 6 studies were included in the meta-analysis. Their combined sensitivity was 98% (95% CI 96%-99%), specificity was 98% (95% CI 97%-99%), LDOR was 7.99 (95% CI 6.73-9.31), and AUC was 0.99 (95% CI 0.99-1). The subgroup analysis showed that the diagnostic performance was acceptable, although imbalanced data were still emphasized as a major problem. All studies had at least one domain with a high risk of bias, and for 20% (9/45) of studies, there were no applicability concerns. CONCLUSIONS: This review summarizes recent evidence in automatic multilevel pain estimation from facial expressions and compared the test accuracy of results in a meta-analysis. Promising performance for pain estimation from facial images was established by current CV algorithms. Weaknesses in current studies were also identified, suggesting that larger databases and metrics evaluating multiclass classification performance could improve future studies. TRIAL REGISTRATION: PROSPERO CRD42023418181; https://www.crd.york.ac.uk/prospero/display_record.php?RecordID=418181.

Correction: Application of AI in Multilevel Pain Assessment Using Facial Images: Systematic Review and Meta-Analysis.

[This corrects the article DOI: 10.2196/51250.].

Cognitive and Hippocampal Changes in Older Adults With Subjective Cognitive Decline After Acupuncture Intervention.

OBJECTIVE: Converging evidence indicates that subjective cognitive decline (SCD) could be an early indicator of dementia. The hippocampus is the earliest affected region during the progression of cognitive impairment. However, little is known about whether and how acupuncture change the hippocampal structure and function of SCD individuals. METHODS: Here, we used multi-modal MRI to reveal the mechanism of acupuncture in treating SCD. Seventy-two older participants were randomized into acupuncture or sham acupuncture group and treated for 12 weeks. RESULTS: At the end of the intervention, compared to sham acupuncture, participants with acupuncture treatment showed improvement in composite Z score from multi-domain neuropsychological tests, as well as increased hippocampal volume and functional connectivity. Moreover, the greater white matter integrity of the fornix, which is the major output tract of the hippocampus, was shown in the acupuncture group. CONCLUSION: These findings suggest that acupuncture may improve the cognitive function of SCD individuals, and increase hippocampal volume on the regional level and enhance the structural and functional connectivity of hippocampus on the connective level.

Inhalable hybrid nanovaccines with virus-biomimetic structure boost protective immune responses against SARS-CoV-2 variants.

BACKGROUND: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), with different antigenic variants, has posed a significant threat to public health. It is urgent to develop inhalable vaccines, instead of injectable vaccines, to elicit mucosal immunity against respiratory viral infections. METHODS: We reported an inhalable hybrid nanovaccine (NVRBD-MLipo) to boost protective immunity against SARS-CoV-2 infection. Nanovesicles derived from genetically engineered 293T cells expressing RBD (NVRBD) were fused with pulmonary surfactant (PS)-biomimetic liposomes containing MPLA (MLipo) to yield NVRBD-MLipo, which possessed virus-biomimetic structure, inherited RBD expression and versatile properties. RESULTS: In contrast to subcutaneous vaccination, NVRBD-MLipo, via inhalable vaccination, could efficiently enter the alveolar macrophages (AMs) to elicit AMs activation through MPLA-activated TLR4/NF-κB signaling pathway. Moreover, NVRBD-MLipo induced T and B cells activation, and high level of RBD-specific IgG and secretory IgA (sIgA), thus elevating protective mucosal and systemic immune responses, while reducing side effects. NVRBD-MLipo also demonstrated broad-spectrum neutralization activity against SARS-CoV-2 (WT, Delta, Omicron) pseudovirus, and protected immunized mice against WT pseudovirus infection. CONCLUSIONS: This inhalable NVRBD-MLipo, as an effective and safe nanovaccine, holds huge potential to provoke robust mucosal immunity, and might be a promising vaccine candidate to combat respiratory infectious diseases, including COVID-19 and influenza.

Dauricine attenuates Oct4/sonic hedgehog co-activated stemness and induces reactive oxygen species-mediated mitochondrial apoptosis via AKT/ß-catenin signaling in human neuroblastoma and glioblastoma stem-like cells.

Neuroblastoma and glioblastoma are primary malignant tumors of the nervous system, with frequent relapse and limited clinical therapeutic drugs. The failure of their treatment is due to the tumor cells exhibiting cancer stem-like cells (CSLCs) properties. Octamer binding transcription factor 4 (Oct4) is involved in mediating CSLCs, our previous work found that Oct4-driven reprogramming of astrocytes into induced neural stem cells was potentiated with continuous sonic hedgehog (Shh) stimulation. In this study, we aimed to study the importance of Oct4 and Shh combination in the stemness properties induction of neuroblastoma and glioblastoma cells, and evaluate the anti-stemness effect of dauricine (DAU), a natural product of bis-benzylisoquinoline alkaloid. The effect of Oct4 and Shh co-activation on cancer stemness was evaluated by tumor spheres formation model and flow cytometry analysis. Then the effects of DAU on SH-SY5Y and T98-G cells were assessed by the MTT, colony formation, and tumor spheres formation model. DAU acts on Oct4 were verified using the Western blotting, MTT, and so on. Mechanistic studies were explored by siRNA transfection assay, Western blotting, and flow cytometry analysis. We identified that Shh effectively improved Oct4-mediated generation of stemness in SH-SY5Y and T98-G cells, and Oct4 and Shh co-activation promoted cell growth, the resistance of apoptosis. In addition, DAU, a natural product, was found to be able to attenuate Oct4/Shh co-activated stemness and induce cell cycle arrest and apoptosis via blocking AKT/ß-catenin signaling in neuroblastoma and glioblastoma, which contributed to the neuroblastoma and glioblastoma cells growth inhibition by DAU. In summary, our results indicated that the treatment of DAU may be served as a potential therapeutic method in neuroblastoma and glioblastoma.

Leveraging a KRAS-based signature to predict the prognosis and drug sensitivity of colon cancer and identifying SPINK4 as a new biomarker.

KRAS is one of the leading mutations reported in colon cancer. However, there are few studies on the application of KRAS related signature in predicting prognosis and drug sensitivity of colon cancer patient. We identified KRAS related differentially expressed genes (DEGs) using The Cancer Genome Atlas (TCGA) database. A signature closely related to overall survival was recognized with Kaplan-Meier survival analysis and univariate cox regression analysis. Then we validated this signature with overall expression score (OE score) algorithm using both scRNA-seq and bulk RNA-seq data. Based on this signature, we performed LASSO cox regression to establish a prognostic model, and corresponding scores were calculated. Differences in genomic alteration, immune microenvironment, drug sensitivity between high- and low-KRD score groups were investigated. A KRAS related signature composed of 80 DEGs in colon cancer were recognized, among which 19 genes were selected to construct a prognostic model. This KRAS related signature was significantly correlated with worse prognosis. Furthermore, patients who scored lower in the prognostic model presented a higher likelihood of responding to chemotherapy, targeted therapy and immunotherapy. Furthermore, among the 19 selected genes in the model, SPINK4 was identified as an independent prognostic biomarker. Further validation in vitro indicated the knockdown of SPINK4 promoted the proliferation and migration of SW48 cells. In conclusion, a novel KRAS related signature was identified and validated based on clinical and genomic information from TCGA and GEO databases. The signature was proved to regulate genomic alteration, immune microenvironment and drug sensitivity in colon cancer, and thus might serve as a predictor for individual prognosis and treatment.

Cantharidin induces apoptosis of human triple negative breast cancer cells through mir-607-mediated downregulation of EGFR.

BACKGROUND: Triple negative breast cancer (TNBC) is a major subtype of breast cancer, with limited therapeutic drugs in clinical. Epidermal growth factor receptor (EGFR) is reported to be overexpressed in various TNBC cells. Cantharidin is an effective ingredient in many clinical traditional Chinese medicine preparations, such as Delisheng injection, Aidi injection, Disodium cantharidinate and vitamin B6 injection. Previous studies showed that cantharidin had satisfactory pharmacological activity on a variety of tumors. In this study, we aimed to study the therapeutic potential of cantharidin for TNBC treatment by targeting EGFR, and expound its novel regulator miR-607. METHODS: The effect of cantharidin on breast cancer in vivo was evaluated by 4T1 mice model. Then the effects of cantharidin on TNBC cells was assessed by the MTT, colony formation, and AnnexinV-PE/7AAD staining. Cantharidin acts on EGFR were verified using the cell membrane chromatography, RT-PCR, Western blotting, MTT, and so on. Mechanistic studies were explored by dual-luciferase report assay, RT-PCR, western blotting, and immunofluorescence staining assay. RESULTS: Cantharidin inhibited TNBC cell growth and induce apoptosis by targeting EGFR. miR-607 was a novel EGFR regulator and exhibited suppressive functions on TNBC cell behaviors. Mechanistic study showed that cantharidin blocked the downstream PI3K/AKT/mTOR and ERK/MAPK signaling pathway. CONCLUSION: Our results revealed that cantharidin may be served as a potential candidate for TNBC treatment by miR-607-mediated downregulation of EGFR.

Caveolin-3 negatively regulates endocytic recycling of cardiac KATP channels.

Sarcolemmal ATP-sensitive potassium (KATP) channels play a vital role in cardioprotection. Cardiac KATP channels are enriched in caveolae and physically interact with the caveolae structural protein caveolin-3 (Cav3). Disrupting caveolae impairs the regulation of KATP channels through several signaling pathways. However, the direct functional effect of Cav3 on KATP channels is still poorly understood. Here, we used the cardiac KATP channel subtype, Kir6.2/SUR2A, and showed that Cav3 greatly reduced KATP channel surface density and current amplitude in a caveolae-independent manner. A screen of Cav3 functional domains revealed that a 25 amino acid region in the membrane attachment domain of Cav3 is the minimal effective segment (MAD1). The peptide corresponding to the MAD1 segment decreased KATP channel current in a concentration-dependent manner with an IC50 of ∼5 µM. The MAD1 segment prevented KATP channel recycling, thus decreasing KATP channel surface density and abolishing the cardioprotective effect of ischemic preconditioning. Our research identified the Cav3 MAD1 segment as a novel negative regulator of KATP channel recycling, providing pharmacological potential in the treatment of diseases with KATP channel trafficking defects.NEW & NOTEWORTHY Cardiac KATP channels physically interact with caveolin-3 in caveolae. In this study, we investigated the functional effect of caveolin-3 on KATP channel activity and identified a novel segment (MAD1) in the C-terminus domain of Caveolin-3 that negatively regulates KATP channel surface density and current amplitude by impairing KATP channel recycling. The peptide corresponding to the MAD1 segment abolished the cardioprotective effect of ischemic preconditioning.

Upper trapezius muscle elasticity in cervical myofascial pain syndrome measured using real-time ultrasound shear-wave elastography.

Background: Myofascial pain syndrome (MPS) is a common cause of neck pain, which is a global public health problem. Because MPS does not present morphological changes within lesioned muscles, there are no imaging diagnostic criteria for this condition. In this study, we evaluate elasticity changes in upper trapezius muscles most frequently involved in cervical MPS using real-time ultrasound shear-wave elastography, and we examine their potential diagnostic value. Methods: We consecutively enrolled 109 right posterior neck pain patients for this prospective study. Of these, 51 were diagnosed with MPS and 58 with non-MPS in the right side of their neck. Among MPS patients, 19 fell into the 1-3 range (mild pain) for pain scores on the visual analog scale (VAS), 25 fell into the 4-6 range (moderate pain), and 7 into the 7-10 range (severe pain). MPS was diagnosed by two independent clinicians using the diagnostic criteria proposed by Simons et al. Using real-time ultrasound shear-wave elastography, we measured right trapezius mean shear-wave velocity (SWVmean). The midpoint of the line between the foramen magnum and the end of the right acromion served as measuring point. Regions of interest were scaled to span 0-8.0 m/s. Results: Trapezius SWVmean was significantly higher in MPS patients compared with non-MPS patients (P<0.001). Stratified analysis of MPS patients according to pain severity revealed similar trapezius SWVmean between mild pain and non-MPS patients (P=0.324), however SWVmean was higher in moderate and severe pain MPS patients compared with non-MPS patients (P<0.001). The area under the curve (AUC) value for upper trapezius SWVmean in MPS patients was 0.791 (95% CI: 0.703-0.863). Corresponding sensitivity and specificity values were 86.27% (95% CI: 73.7-94.3%) and 62.07% (95% CI: 48.4-74.5%). Stratified analysis of MPS patients by pain severity produced the following AUC values for trapezius SWVmean in MPS patients with mild, moderate, and severe pain: 0.578 (95% CI: 0.460-0.690), 0.899 (95% CI: 0.814-0.955), and 0.983 (95% CI: 0.914-0.999), respectively. Conclusions: Elasticity changes and increased stiffness in the trapezius occur in cervical MPS patients with moderate and severe pain. The SWVmean parameter reflecting trapezius muscle elasticity may be valuable for successful screening of cervical MPS, especially in patients with moderate and severe pain.

Multi-Site Attack, Neutrophil Membrane-Camouflaged Nanomedicine with High Drug Loading for Enhanced Cancer Therapy and Metastasis Inhibition.

Background: Advanced breast cancer is a highly metastatic tumor with high mortality. Simultaneous elimination of primary tumor and inhibition of neutrophil-circulation tumor cells (CTCs) cluster formation are urgent issues for cancer therapy. Unfortunately, the drug delivery efficiency to tumors and anti-metastasis efficacy of nanomedicine are far from satisfactory. Methods: To address these problems, we designed a multi-site attack, neutrophil membrane-camouflaged nanoplatform encapsulating hypoxia-responsive dimeric prodrug hQ-MMAE2 (hQNM-PLGA) for enhanced cancer and anti-metastasis therapy. Results: Encouraged by the natural tendency of neutrophils to inflammatory tumor sites, hQNM-PLGA nanoparticles (NPs) could target delivery of drug to tumor, and the acute hypoxic environment of advanced 4T1 breast tumor promoted hQ-MMAE2 degradation to release MMAE, thus eliminating the primary tumor cells to achieve remarkable anticancer efficacy. Alternatively, NM-PLGA NPs inherited the similar adhesion proteins of neutrophils so that NPs could compete with neutrophils to interrupt the formation of neutrophil-CTC clusters, leading to a reduction in extravasation of CTCs and inhibition of tumor metastasis. The in vivo results further revealed that hQNM-PLGA NPs possessed a perfect safety and ability to inhibit tumor growth and spontaneous lung metastasis. Conclusion: This study demonstrates the multi-site attack strategy provides a prospective avenue with the potential to improve anticancer and anti-metastasis therapeutic efficacy.

STING activation in macrophages by vanillic acid exhibits antineoplastic potential.

The host stimulator of interferon genes (STING) signaling pathway is a major innate immune sensing pathway, and the stimulation of this pathway within antigen-presenting cells shows promise in targeting immune-suppressed tumors. Macrophages resident in tumors exhibit anti-inflammatory properties and enhance tumor growth and development. Polarizing such macrophages towards a pro-inflammatory phenotype is an effective strategy for tumor suppression. In the present study, we observed that the STING pathway was inactivated in breast and lung carcinomas, and a positive correlation existed between STING and macrophage markers in these tumors. We found that vanillic acid (VA) could stimulate the STING/TBK1/IRF3 pathway. VA mediated the production of type I IFN and promoted macrophage polarization into the M1 phenotype; this activity was dependent on STING activation. A direct-contact co-culture model and a transwell co-culture model revealed that macrophages with VA-induced STING activation exhibited anti-proliferative effects on SKBR3 and H1299 cells, although a STING antagonist and M2 macrophage-related cytokines alleviated this anti-proliferative effect. Further investigation indicated that phagocytosis and apoptosis-inducing effects were the major mediators of the anti-tumor effect of VA-treated macrophages. Mechanistically, VA promoted the polarization of macrophages to a M1 phenotype via IL-6R/JAK signaling, resulting in enhanced phagocytosis and apoptosis-induction effects. Additionally, STING activation-induced IFNß production also participated in the apoptosis mediated by VA-treated macrophage in SKBR3 and H1299 cells. Mouse models with 4 T1 tumors confirmed the anti-tumor properties of VA in vivo and revealed the infiltration of VA-induced cytotoxic T cells into the tumors. These data suggest that VA is an effective agonist of STING and provides a new perspective for cancer immunotherapy.

IL-17A-mediated mitochondrial dysfunction induces pyroptosis in colorectal cancer cells and promotes CD8 + T-cell tumour infiltration.

BACKGROUND: Interleukin-17A (IL-17A), a proinflammatory cytokine primarily secreted by Th17 cells, γδT cells and natural killer T (NKT) cells, performs essential roles in the microenvironment of certain inflammation-related tumours by regulating cancer growth and tumour elimination proved in previous literature. In this study, the mechanism of IL-17A that induces mitochondrial dysfunction promoted pyroptosis has been explored in colorectal cancer cells. METHOD: The records of 78 patients diagnosed with CRC were reviewed via the public database to evaluate clinicopathological parameters and prognosis associations of IL-17A expression. The colorectal cancer cells were treated with IL-17A, and the morphological characteristics of those cells were indicated by scanning electron microscope and transmission electron microscope. After IL-17A treatment, mitochondrial dysfunction was tested by mitochondrial membrane potential (MMP) and reactive oxygen species (ROS). The expression of pyroptosis associated proteins including cleaved caspase-4, cleaved gasdermin-D (GSDMD), IL-1ß, receptor activator of nuclear NOD-like receptor family pyrin domain containing 3 (NLRP3), apoptosis-associated speck like protein containing a card (ASC), and factor-kappa B was measured through western blotting. RESULTS: Positive IL-17A protein expression was observed in CRC compared to the non-tumour tissue. IL-17A expression indicates a better differentiation, earlier stage, and better overall survival in CRC. IL-17A treatment could induce mitochondrial dysfunction and stimulate intracellular reactive oxygen species (ROS) production. Furthermore, IL-17A could promote pyroptosis of colorectal cancer cells and significantly increase the secretion of inflammatory factors. Nevertheless, the pyroptosis induced by IL-17A could be inhibited through the pre-treatment with Mito-TEMPO (a mitochondria-targeted superoxide dismutase mimetic with superoxide and alkyl radical scavenging properties) or Z-LEVD-FMK (caspase-4 inhibitor, fluoromethylketone). Additionally, after being treated with IL-17A, an increasing number of CD8 + T cells showed in mouse-derived allograft colon cancer models. CONCLUSION: IL-17A, as a cytokine mainly secreted by γδT cells in the colorectal tumour immune microenvironment, can regulate the tumour microenvironment in multiple ways. IL-17A could induce mitochondrial dysfunction and pyroptosis through the ROS/NLRP3/caspase-4/GSDMD pathway, and promote intracellular ROS accumulation. In addition, IL-17A can promote the secretion of inflammatory factors such as IL-1ß、IL-18 and immune antigens, and recruit CD8 + T cells to infiltrate tumours.

Near-infrared magnetic core-shell nanoparticles based on lanthanide metal-organic frameworks as a ratiometric felodipine sensing platform.

Felodipine is an effective drug to treat hypertension, but its abuse can cause bardycardia. It is significant to develop highly sensitive detection platform for felodipine to enable the efficient treatment of hypertension diseases. In this work, to highly efficiently detect felodipine, multi-emission near-infrared (NIR) hierarchical magnetic core-shell lanthanide-MOF nanoparticles, namely Nd-MOF@Yb-MOF@SiO2@Fe3O4 (NIR-1), has been synthesized by layer-by-layer (LBL) method. LBL method can adjust the optical properties of NIR-1 and expose more active sites to improve sensitivity in detection process. NIR-1 has near-infrared luminescence emission, which can efficiently avoid the interference of autofluorescence in biological tissues. Photo-luminescent (PL) experiments also reveal that NIR-1 could be used as a near-infrared ratiometric luminescent sensor for felodipine detection with high selectivity and sensitivity, the low of detection limit (LOD) is 6.39 nM in felodipine detection, which is also performed using real biological samples. In addition, NIR-1 can be used as a ratiometric thermometer could also be applied in the temperature sensing from 293 K to 343 K. Finally, detection mechanisms for felodipine and temperature sensing performance based on near-infrared (NIR) emission were also investigated and discussed in detail.

["Houxi (SI 3) communicating the governor vessel" verified with the infrared thermal imaging technology].

OBJECTIVE: To compare the effect on facial acupoint temperature between acupuncture at Houxi (SI 3) and Dazhui (GV 14) so as to verify "Houxi (SI 3) communicating the governor vessel" based on the infrared thermal imaging technology. METHODS: Thirty-five healthy subjects (5 cases dropped off) were collected and before-after study in the same subject was adopted. The subjects were successively assigned into a sham-acupuncture group, a Houxi group, a Wangu group and a Dazhui group. Sham-acupuncture at Houxi (SI 3) on the left, acupuncture at Houxi (SI 3) on the left, Wangu (SI 4) on the left and Dazhui (GV 14) were given respectively. One intervention was given and the needles were retained for 30 min in each group. 30 min before and after acupuncture, the infrared thermal images of the face were collected, and the facial temperature was compared among the following 5 acupoints, i.e. Yintang (GV 24+), Suliao (GV 25), Shuigou (GV 26), Duiduan (GV 27) and Chengjiang (CV 24). RESULTS: After acupuncture, the facial temperature at Yintang (GV 24+) and Chengjiang (CV 24) was increased compared before acupuncture in the sham-acupuncture group (P<0.01, P<0.05). The facial temperature at Suliao (GV 25) in the Houxi group was reduced after acupuncture (P<0.05). In the Wangu group, the temperature at Yintang (GV 24+) was increased compared before acupuncture (P<0.01). The facial temperature was increased at Duiduan (GV 27) and Chengjiang (CV 24) compared before acupuncture in the Dazhui group (P<0.01, P<0.05). The differences of facial temperature at Chengjiang (CV 24) and Suliao (GV 25) after acupuncture were larger than before acupuncture in the Houxi group and the Dazhui group (P<0.01). In comparison with the temperature at Suliao (GV 25) of the same group, the differences of facial temperature before and after acupuncture at Yintang (GV 24+), Shuigou (GV 26), Duiduan (GV 27) and Chengjiang (CV 24) were increased in the Houxi group (P<0.01, P<0.05); while, the increase was also obtained at Yintang (GV 24+), Shuigou (GV 26), Duiduan (GV 27) and Chengjiang (CV 24) in the Dazhui group (P<0.05, P<0.01). The difference of facial temperature at Yintang (GV 24+) before and after acupuncture was increased compared with Suliao (GV 25) in the Wangu group (P<0.05). CONCLUSION: Acupuncture at Houxi (SI 3) generates a similar thermal effect as Dazhui (GV 14). It regulates and dissipates the core temperature to "govern the yang qi of the whole body".

The cardioprotective role of sirtuins is mediated in part by regulating KATP channel surface expression.

Sirtuins are NAD+-dependent deacetylases with beneficial roles in conditions relevant to human health, including metabolic disease, type II diabetes, obesity, cancer, aging, neurodegenerative diseases, and cardiac ischemia. Since ATP-sensitive K+ (KATP) channels have cardioprotective roles, we investigated whether they are regulated by sirtuins. Nicotinamide mononucleotide (NMN) was used to increase cytosolic NAD+ levels and to activate sirtuins in cell lines, isolated rat and mouse cardiomyocytes or insulin-secreting INS-1 cells. KATP channels were studied with patch clamping, biochemistry techniques, and antibody uptake experiments. NMN led to an increase in intracellular NAD+ levels and an increase in the KATP channel current, without significant changes in the unitary current amplitude or open probability. An increased surface expression was confirmed using surface biotinylation approaches. The rate of KATP channel internalization was diminished by NMN, which may be a partial explanation for the increased surface expression. We show that NMN acts via sirtuins since the increased KATP channel surface expression was prevented by blockers of SIRT1 and SIRT2 (Ex527 and AGK2) and mimicked by SIRT1 activation (SRT1720). The pathophysiological relevance of this finding was studied using a cardioprotection assay with isolated ventricular myocytes, in which NMN protected against simulated ischemia or hypoxia in a KATP channel-dependent manner. Overall, our data draw a link between intracellular NAD+, sirtuin activation, KATP channel surface expression, and cardiac protection against ischemic damage.

Imaging Features and Risk Factors of Pancreatic Cystic Lesions Complicating Autoimmune Pancreatitis: A Retrospective Study.

OBJECTIVE: This study aimed to explore the imaging features and risk factors of PCLs complicating AIP, and investigate its prognosis through continuous imaging follow-up. PATIENTS AND METHODS: Patients who were diagnosed with AIP from January 2014 to December 2020 in our hospital were recruited. We analyzed the CT and MRI features of PCLs complicating AIP, and investigated its prognosis through imaging follow-up. We also compared subjects with and without PCLs using clinical, laboratory, and imaging data; the related risk factors associated with PCLs were investigated in a multivariate logistic regression analysis. RESULTS: In this group, 16 patients had PCLs and 86 did not. A total of 43 PCLs larger than 5mm were found in 15 patients. Among these PCLs, 35 showed homogeneous signal (density); one, bleeding; three, linear separation; and four, small focal low signal on T2WI. Eight patients with 23 PCLs appeared for the follow-up after steroid treatment. Short-term follow-up showed that 11 PCLs disappeared, nine reduced, one unchanged and two enlarged. Of the 12 PCLs that did not disappear, 10 PCLs disappeared at long-term follow-up, except for two reduced PCLs were not re-examined. Logistic regression analysis showed that drinking history was an independent risk factor, age ≥ 65 years was an independent protective factor for PCLs complicating AIP. CONCLUSION: The imaging features of PCLs complicating AIP are various, which can be single or multiple, most of them are homogeneous, and some lesions may be accompanied by hemorrhage, separation and necrosis. Age ≥ 65 years and avoiding drinking may help to reduce the occurrence of these lesions.

Circ_C4orf36 Promotes the Proliferation and Osteogenic Differentiation of BMSCs by Regulating VEGFA.

Fracture healing is a complicated process containing the regulation of cellular process. It has been reported that circRNAs are involved in fracture healing. Our study aims to explore the role and mechanism of circ_C4orf36 in fracture healing. Here, we found that the expressions of Circ_C4orf36 and VEGFA were increased during osteoblast differentiation in MC3T3-E1 cells. Circ_C4orf36 overexpression could accelerate the proliferation and migration, as well as osteoblast differentiation in MC3T3-E1 cells, as well as increased ALP activity and osteogenic markers (Runx2, OCN) via upregulating VEGFA expression. Mechanistically, circ_C4orf36 facilitated the expression of VEGFA by recruiting EIF4A3. Taken together, our results elucidated that circ_C4orf6 promoted the migration, proliferation and osteoblast differentiation in BMSCs by upregulating VEGFA, which indicated that circ_C4orf36 might be a potential target in fracture healing treatment.

Splenic-targeting biomimetic nanovaccine for elevating protective immunity against virus infection.

BACKGROUND: The prevalence of viral infectious diseases has become a serious threat to public safety, economic and social development. Vaccines have been served as the most effective platform to prevent virus transmission via the activation of host immune responses, while the low immunogenicity or safety, the high cost of production, storage, transport limit their effective clinical application. Therefore, there is a need to develop a promising strategy to improve the immunogenicity and safety of vaccines. METHODS: We developed a splenic-targeting biomimetic nanovaccine (NV) that can boost protective humoral and cellular immunity against african swine fever virus (ASFV) infection. The universal PLGA nanoparticles (CMR-PLGA/p54 NPs) coated with mannose and CpG (TLR9 agonist) co-modified red blood cell (RBC) membrane were prepared, which comprised a viral antigen (p54) and can be served as a versatile nanovaccine for elevating protective immunity. RESULTS: CMR-PLGA/p54 NVs could be effectively uptaken by BMDC and promoted BMDC maturation in vitro. After subcutaneous immunization, antigen could be effectively delivered to the splenic dendritic cells (DCs) due to the splenic homing ability of RBC and DC targeting capacity of mannose, which promoted antigen presentation and DCs maturation, and further elicited higher levels of cytokines secretion and specific IgG titers, CD4+ and CD8+ T cells activation and B maturation. Moreover, NVs demonstrated notable safety during the immunization period. CONCLUSIONS: This study demonstrates the high potential of CMR-PLGA NPs as vaccine delivery carriers to promote humoral and cellular immune responses, and it provides a promising strategy to develop safe and effective vaccines against viral infectious diseases.