The Janus kinase/signal transducers and activators of transcription (JAK-STAT) control natural killer (NK) cells development and cytotoxic functions, however, whether long non-coding RNAs (lncRNAs) are involved in this pathway remains unknown. We found that miR155HG was elevated in activated NK cells and promoted their proliferation and effector functions in both NK92 and induced-pluripotent stem cells (iPSCs)-derived NK (iPSC-NK) cells, without reliance on its derived miR-155 and micropeptide P155. Mechanistically, miR155HG bound to miR-6756 and relieved its repression of JAK3 expression, thereby promoting the JAK-STAT pathway and enhancing NK cell proliferation and function. Further investigations disclosed that upon cytokine stimulation, STAT3 directly interacts with miR155HG promoter and induces miR155HG transcription. Collectively, we identify a miR155HG-mediated positive feedback loop of the JAK-STAT signaling. Our study will also provide a power target regarding miR155HG for improving NK cell generation and effector function in the field of NK cell adoptive transfer therapy against cancer, especially iPSC-derived NK cells.

This study investigated the regulatory potential of salidroside (SAL), a primary active compound in Rhodiola rosea L., on osteoclast differentiation by modulating the hypoxia-inducible factor 1-alpha (HIF-1a) pathway in osteoblasts. Luciferase reporter assay and chromatin immunoprecipitation (ChIP) assay were employed to validate whether the receptor activator of nuclear factor-?B ligand (RANKL) is the downstream target gene of HIF-1a in osteoblasts. The study also utilized lipopolysaccharide (LPS)-induced mouse osteolysis to examine the impact of SAL on osteolysis in vivo. Furthermore, conditioned medium (CM) from SAL-pretreated osteoblasts was used to investigate the paracrine effects on osteoclastogenesis through the HIF-1a pathway. Hypoxic condition-induced overexpression of HIF-1a upregulated RANKL levels by binding to the RANKL promoter and enhancing transcription in osteoblastic cells. In vivo, SAL significantly alleviated bone tissue hypoxia and decreased the expression of HIF-1a by downregulating the expression of RANKL, vascular endothelial growth factor (VEGF), interleukin 6 (IL-6), and angiopoietin-like 4 (ANGPTL4). In the paracrine experiment, conditioned media from SAL-pretreated osteoblasts inhibited differentiation through the HIF-1a/RANKL, VEGF, IL-6, and ANGPTL4 pathways. RANKL emerges as the downstream target gene regulated by HIF-1a in osteoblasts. SAL significantly alleviates bone tissue hypoxia and bone loss in LPS-induced osteolysis through the HIF-1a/RANKL, VEGF, IL-6, and ANGPTL4 pathways. SAL inhibits osteoclast differentiation by regulating osteoblast paracrine secretion.
Animals ; Osteoblasts/cytology* ; Hypoxia-Inducible Factor 1, alpha Subunit/genetics* ; Glucosides/administration & dosage* ; Cell Differentiation/drug effects* ; Phenols/administration & dosage* ; Mice ; Osteoclasts/metabolism* ; RANK Ligand/genetics* ; Rhodiola/chemistry* ; Osteogenesis/drug effects* ; Signal Transduction/drug effects* ; Interleukin-6/genetics* ; Male ; RAW 264.7 Cells ; Osteolysis/genetics* ; Humans ; Mice, Inbred C57BL

Objective To investigate the relationship between polymorphism of resistin(RETN)gene and metabolic associated fatty liver disease(MAFLD)in type 2 diabetes mellitus(T2DM)patients in middle and high altitude areas.Methods A total of 400 patients with T2DM in Qinghai area were recruited and divided into simple T2DM group(T2DM,n=200)and T2DM combined with MAFLD group(T2DM+ MAFLD,n=200)according to liver ultrasonography.Healthy individuals confirmed by physical examination were selected as the normal control group(NC,n=180).Plasma resistin levels were measured by ELISA.The polymorphism of RETN-420C/G and +299G/A genes were detected by PCR sequencing.Results By comparing the polymorphism of RETN-420C/G gene in each group,it was found that the frequencies of G/G genotype and G allele frequency in T2DM+MAFLD group were higher than those in NC group and T2DM group(P<0.05),while the frequencies of C/C genotype and C allele frequency were lower than those in NC group and T2DM group(P<0.05).The risk of MAFLD increased by 1.571,2.126 and 1.537 times respectively in T2DM patients with C/G,G/G genotype and G allele.Logistic regression analysis showed that G/G genotype was a risk factor for MAFLD in T2DM patients.By comparing the polymorphism of RETN+299G/A gene in each group,it was found that A allele frequency in T2DM+MAFLD group was higher than that in NC group and T2DM group,while G allele frequency was lower than that in NC group and T2DM group(P<0.05).The allele A increased the risk of MAFLD in T2DM patients by 1.432 times compared to allele G.Conclusion RETN gene-420C/G locus G/G genotype increases the risk of T2DM combined with MAFLD in middle and high altitudeareas.

Objective To analyze the effect of neurofeedback training based on early start Denver model(ESDM)on children with autism spectrum disorder(ASD). Methods From August,2020 to May,2024,a total of 60 children with ASD from Hangzhou Children's Hospital were randomly divided into control group(n=30)and observation group(n=30).The control group received ESDM intervention,while the observation group received neurofeedback training in addition,for six months.They were assessed with Autism Treatment Evaluation Checklist(ATEC)and Psycho-Educational Profile-3rd Edition(PEP-3). Results After treatment,the score of ATEC was lower in the observation group than in the control group(t=3.545,P<0.05),the scores of cognition(t=2.236,P=0.029),emotional expression(t=2.293,P=0.025)and problem be-havior(Z=2.099,P=0.036)were higher in the observation group than in the control group.The score differenc-es of ATEC(Z=3.620,P<0.001),and cognition(Z=2.920,P<0.05)and problem behaviors(Z=4.209,P<0.05)of PEP-3 before and after intervention were higher in the observation group than in the control group. Conclusion Combination of neurofeedback training could improve the effect of ESDM on ASD.

ObjectiveTo investigate the attenuating effect of Dioscoreae Bulbiferae Rhizoma（DBR） processed with Phaseoli Radiati Semen（PRS） juice， and explore the attenuating mechanism based on ferroptosis of the main toxic target organ. MethodSixty male ICR mice were randomly divided into blank group， DBR group， water roasted DBR group（hereinafter referred to as water group）， PRS juice-roasted DBR group 1（DBR-PRS 10∶1， stuffy moistening for 40 min， stir-fried at 130 ℃ for 18 min， hereinafter referred to as group 1）， PRS juice-roasted DBR group 2（DBR-PRS 10∶1， stuffy moistening for 80 min， stir-fried at 100 ℃ for 14 min， hereinafter referred to as group 2）， PRS juice-roasted DBR group 3（DBR-PRS=20∶3， stuffy moistening for 40 min， stir-fried at 160 ℃ for 14 min， hereinafter referred to as group 3）. The raw and processed groups of DBR were gavaged with their corresponding 95% ethanol extract at a dose of 3 g·kg^-1·d^-1， while the blank group was gavaged with an equal volume of 0.5% sodium carboxymethyl cellulose， once a day for 14 consecutive days. Hematoxylin-eosin（HE） staining was used to observe the histopathological changes of mouse liver. Alanine aminotransferase（ALT） and aspartate aminotransferase（AST） levels in serum， as well as malondialdehyde（MDA）， ferrous ions（Fe²⁺）， reduced glutathione（GSH） and superoxide dismutase（SOD） levels in liver tissue were detected by the biochemical detection. Western blot was used to detect the expression of iron key proteins such as ferritin heavy chain 1（FTH1） and glutathione peroxidase 4（GPX4）. ResultHE staining results showed that the liver tissue structure of the blank group was clear， the morphology of hepatocytes was normal， the cytoplasms of hepatocytes in the DBR group and water group were loose and vacuolar， with obvious pathological damages， and the pathologic damages of mice in the group 1-3 were significantly improved. Compared with the blank group， the levels of ALT， AST， MDA and Fe²⁺ in mice from the DBR group were significantly increased（P<0.01）， while GSH and SOD levels were significantly reduced（P<0.01）， and the protein expression levels of FTH1 and GPX4 were significantly decreased（P<0.01）. Compared with the DBR group， the ALT， AST，MDA and Fe²⁺ levels of mice in the group 1-3 were significantly reduced（P<0.05， P<0.01）， the GSH and SOD levels and the protein expression levels of FTH1 and GPX4 were significantly increased（P<0.01）. Compared with the water group， the AST and MDA levels of mice in the group 1-3 were significantly reduced（P<0.05， P<0.01）， the SOD level significantly increased（P<0.05， P<0.01）， the FTH1 protein expression significantly increased（P<0.01）， and the serum ALT level of mice in the group 2-3 significantly reduce（P<0.01）， Fe²⁺ level significantly reduced（P<0.01）， GSH level significantly increased（P<0.05， P<0.01）， and GPX4 protein expression significantly increased（P<0.05， P<0.01）. Among the group 1-3， the group 3 had the best detoxification effect. ConclutionProcessing with PRS juice can reduce the liver injury induced by DBR， and the mechanism may be related to the inhibition of ferroptosis in the liver.

Objective:To assess the prognostic impact of frailty on elderly inpatients with heart failure.Methods:This prospective cohort study enrolled 121 in elderly patients with heart failure from Beijing Hospital, the General Hospital of the People's Liberation Army, and Beijing Tsinghua Changgung Hospital between September 2018 and April 2019.Patients were assessed for frailty using the Fried frailty phenotype and categorized into frail and non-frail groups.Follow-ups were conducted at 3-, 6-, and 12-months post-enrollment through clinic visits or phone calls to record adverse events.Composite endpoints include all-cause mortality and rehospitalization duo to deterioration of heart failure.Results:The study included 121 patients with an average age of 78.0±7.4 years, of whom 71(58.7%)were male and 57(47.1%)were classified as frail.Compared to the non-frail group, the frail group had lower estimated glomerular filtration rates[49.5±20.7 ml/(min·1.73m 2) vs.(64.0±27.1)ml/(min·1.73m 2)], lower scores in Basic Activities of Daily Living[5.0(4.0, 6.0) vs.6.0(5.0, 6.0)], Instrumental Activities of Daily Living[2.0(1.3, 7.8) vs.7.0(5.0, 8.0)], and Mini-Mental State Examination[26.0(16.0, 28.0) vs.27.0(22.3, 29.0)], all P<0.05.They also experienced longer hospital stays[10.5(6.0, 18.8)days vs.8.0(6.0, 11.8)days, P=0.008].During the follow-up period, the incidence of composite endpoint events was significantly higher in the frail group(43.9% vs.25.0%, P=0.029).Kaplan-Meier survival analysis demonstrated that the one-year incidence of composite endpoint events was significantly higher in the frail group( P=0.013).Multivariable Cox regression analysisindicated that frailty was an independent risk factor for composite endpoint events( HR=2.201, 95% CI: 1.089-4.447, P=0.028). Conclusions:Frailty is an independent risk factor for poor outcomes in elderly hospitalized patients with heart failure and should be considered a crucial factor in clinical assessment and treatment strategies.

Objective:To explore the changes of large-scale functional brain networks and network topological properties in patients with major depressive disorder (MDD) whose diagnosis had not changed after 5 years of follow-up.Methods:Totally 521 cases of hospitalized MDD patients were recruited from January 2012 to August 2018, and another 204 cases of gender- and age-matched healthy controls were recruited. All participants completed resting-state functional magnetic resonance scanning and clinical assessment. Their diagnosis were reviewed 5 years after discharge.A total of 258 participants whose diagnosis had not changed were counted into the MDD group for analysis. The differences in large-scale brain network connectivity between the two groups were analyzed by constructing a whole-brain functional network, on the basis of which the altered topological properties of the sensorimotor network (SMN), visual network (VN) and default mode network (DMN) were further analyzed between the two groups.The SPSS 24.0 software was used for data analysis and the independent sample t-test and χ2 test were used for the data comparison of the two groups. Results:Compared with the healthy controls, the MDD group had significantly decreased network clustering, mainly involving the SMN, VN and DMN (edge P<0.001, cluster P<0.05). The MDD group had decreased functional connectivity(FC) strength within the SMN, VN and DMN networks, the FC strength between the SMN and VN networks, between the frontoparietal network (FPN) and the DAN networks were decreased(all P<0.05, FDR corrected). Graph-theory analysis showed that local efficiency, clustering coefficient, and normalized shortest path length were decreased in the MDD group, node efficiency was decreased in the left ventral medial prefrontal cortex and the middle of the bilateral insula, node centrality was decreased in the middle of the bilateral insula and occipital lobe, and the betweenness was decreased in the middle of the right insula (all P<0.05, FDR corrected). Conclusion:MDD exhibits abnormal network functional connectivity, disruption of network topological properties, diminished optimal information processing, and to some extent reflects the severity of depressive symptoms. The decreased ability of information transfer flow in the insula plays an important role for the functional abnormality of the network.

Objective To clone Rhipicephalus linnaei Boophilin-H2 gene, construct the recombinant expression vector, express the Boophilin-H2 recombinant protein in Escherichia coli, and assess its anticoagulant activity. Methods Specific primers were designed to amplify the Boophilin-H2 gene fragment using cDNA, synthesized from engorged Rhipicephalus linnaei tick RNA through reverse transcription, as a template. The gene fragment was cloned and connected to plasmid pSmart-I, and the recombinant expression vector pSmart-I/Boophilin-H2 was constructed. The recombinant expression vector was verified by double restriction enzyme digestion with BamHⅠand XhoⅠ, transferred into the competent state of Escherichia coli BL21(DE3, and expressed under low-temperature induction with IPTG. The recombinant protein was purified by Ni-NTA Resin, and its expression and purification were detected by 12.5% SDS-PAGE. The femoral venous blood of New Zealand white rabbits was collected by 3.8% sodium citrate blood collection tube, and the upper plasma was centrifugally separated to measure the anticoagulant activity of the recombinant protein using four test plates of in vitro coagulation. Results A 387 bp gene fragment of Boophilin-H2 of Rhipicephalus linnaei was successfully amplified and cloned; the prokaryotic expression vector pSmart-I/Boophilin-H2 was constructed and verified by double enzyme digestion. Following induction with 0.8 mmol/L IPTG for 16 hours in Escherichia coli, SDS-PAGE showed that the recombinant protein was expressed in the supernatant primarily in a soluble form, with the Boophilin-H2 recombinant protein approximately 35 000 in size. The anticoagulant activity assays of the purified recombinant protein Boophilin-H2 showed that the recombinant protein significantly prolongs the activated partial thromboplastin time (APTT) in a concentration-dependent manner, while its effects on thrombin time (TT), prothrombin time (PT), and fibrinogen (FIB) levels were not significant. Conclusions The expression vector for Boophilin-H2 was successfully constructed, and its product exhibited potent APTT anticoagulant activity, implying its role in the intrinsic coagulation pathway, possibly acting upon intrinsic coagulation factors VIII, XI, and XII to inhibit blood coagulation. This study provides a reference and theoretical foundation for further research and development of tick control vaccines and anticoagulant drugs.

Objective:To investigate the expression of long non-coding RNA (lncRNA) MTATP6P1 in melanoma and its effect on the proliferation, migration and invasion of melanoma cells by targeting miRNA-411-5p (miR-411-5p).Methods:A total of 461 samples of melanoma tissues and paracancerous tissues (>2 cm from the tumor margin) were collected from the tumor-associated lncRNA database (TANRIC database updated in July 2021), and the expression of MTATP6P1 was compared between the two groups. The bioinformatics software lncRNA Disease v2.0 was used to predict the possible binding site microRNA (miRNA) of MTATP6P1. Real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) was used to detect the relative expression level of MTATP6P1 in melanoma cells A-375, WM266-4, VMM5A, A2058 and normal human epidermal melanocytes PIG1; and the lowest relative expression level of cells in MTATP6P1 were divided into MTATP6P1 group (transfected with MTATP6P1 overexpression plasmid) and NC group (transfected with blank plasmid). The proliferation ability of A-375 cells was detected by using CCK-8 method; the migration ability of A-375 cells was detected by using scratch test; the invasion ability of A-375 cells was detected by using Transwell assay; the targeting relationship between MTATP6P1 and miR-411-5p was detected by using dual luciferase reporter gene assay; Western blot was used to detect the expression of ERK signaling pathway related proteins in cells.Results:The relative expression levels of MTATP6P1 in melanoma tissues and adjacent tissues were 9.82±0.58 and 11.56±0.16, respectively. The expression level of MTATP6P1 in melanoma tissues was lower than that in paracancerous tissues ( t = 9.56, P = 0.009). The relative expression levels of MTATP6P1 in normal human epidermal melanocyte PIG1 and melanoma cells A-375, WM266-4, VMM5A, and A2058 were 1.01±0.13, 0.12±0.02, 0.66±0.04, 0.39±0.07, 0.49±0.05; the relative expression level of MTATP6P1 in melanoma cells was lower than that in PIG1 cells (all P < 0.05), and then A-375 cells with the lowest relative expression level were taken to perform the subsequent experiments. The relative expression levels of MTATP6P1 in A-375 cells of MTATP6P1 group and NC group were 14.83±1.67 and 1.02±0.30, respectively ( t = 8.13, P < 0.001). After 16, 24, 32, and 40 h of culture, the proliferation ability of the cells in the MTATP6P1 group was lower than that in NC group (all P < 0.05). The scratch healing rates of A-375 cells in MTATP6P1 group and NC group were (26±7)% and (55±4)%, respectively; the scratch healing rate in MTATP6P1 group was lower than that in NC group ( t = 3.48, P = 0.009). The invasive number of A-375 cells in MTATP6P1 group and NC group were (32±12) and (116±17), respectively; the number of invasive cells in MTATP6P1 group was lower than that in NC group ( t = 4.11, P = 0.006). The results of dual luciferase reporter gene assay showed that there was a targeting relationship between MTATP6P1 and miR-411-5p. The relative expression level of miR-411-5p in A-375 cells of MTATP6P1 group and NC group was 1.04±0.16 and 5.37±0.68, respectively; the expression level of miR-411-5p in MTATP6P1 group was lower than that in NC group ( t = 6.20, P < 0.001). The expressions of ERK signaling pathway proteins p-Ras, p-Raf, p-MEK1, p-RSK, and AP-1 in A-375 cells of MTATP6P1 group were lower than those in NC group. Conclusions:MTATP6P1 inhibits the proliferation, migration and invasion of melanoma A-375 cells through targeting miR-411-5p.

The gluteal fat grafting technique continues to grow in popularity, along with which the safety issue should not be ignored. It drew attentions of researchers that due to the existence of large vessels, fat embolism or fat embolism syndrome occurred more frequently for gluteal fat grafting, which might lead to death. In this situation, researchers performed a series of study including retrospective case series, systematic analysis, questionnaire survey and anatomical study, and summarized a series of safety principles for gluteal fat grafting, such as scanning high-risk patients; staying subcutaneously injection, avoiding intramuscular injection; using cannula size over 4 mm; keeping cannula parallel to gluteus maximus; injecting while withdrawing; and avoiding excessive local pressure. Based on the literature review, this article analyzes and summarizes the safety of gluteal fat grafting.