Glutamate-ammonia ligase (GLUL, also known as glutamine synthetase) is a crucial enzyme that catalyzes ammonium and glutamate into glutamine in the ATP-dependent condensation. Although GLUL plays a critical role in multiple cancers, the expression and function of GLUL in gastric cancer remain unclear. In the present study, we have found that the expression level of GLUL was significantly lower in gastric cancer tissues compared with adjacent normal tissues, and correlated with N stage and TNM stage, and low GLUL expression predicted poor survival for gastric cancer patients. Knockdown of GLUL promoted the growth, migration, invasion and metastasis of gastric cancer cells in vitro and in vivo, and vice versa, which was independent of its enzyme activity. Mechanistically, GLUL competed with β-Catenin to bind to N-Cadherin, increased the stability of N-Cadherin and decreased the stability of β-Catenin by alerting their ubiquitination. Furthermore, there were lower N-Cadherin and higher β-Catenin expression levels in gastric cancer tissues compared with adjacent normal tissues. GLUL protein expression was correlated with that of N-Cadherin, and could be the independent prognostic factor in gastric cancer. Our findings reveal that GLUL stabilizes N-Cadherin by antagonizing β-Catenin to inhibit the progress of gastric cancer.

ObjectiveTo investigate the association between estimated glucose disposal rate （eGDR） and the severity of coronary heart disease. MethodsWe conducted a hospital-based cross-sectional study that included 1258 patients （mean age： 62（53-68） years） who underwent coronary angiography for suspected coronary artery disease （53.9% were male）. Insulin resistance level （IR） was calculated according to eGDR formula： eGDR = 21.158 - （0.09 × WC） - （3.407 × hypertension） - （0.551 × HbA1c） ［hypertension （yes = 1 / no = 0）， HbA1c = HbA1c （%）］. Subjects were grouped according to the eGDR quantile. CAD severity was determined by the number of narrowed vessels： no-obstructive CAD group （all coronary stenosis were<50%， n=704）， Single-vessel CAD group （only one involved major coronary artery stenosis≥50%， n=205）， Multi-vessel CAD group （two or more involved major coronary arteries stenosis≥50%， n=349）； Multivariate logistic regression model was used to analyze the association between eGDR and CAD severity. The linear relationship between eGDR and CAD in the whole range of eGDR was analyzed using restricted cubic spline. Subgroup analyses were used to assess the association between eGDR and CAD severity in different diabetic states. Receiver operating characteristic （ROC） curve analysis were used to evaluate the value of eGDR in improving CAD recognition. ResultsA decrease in the eGDR index was significantly associated with an increased risk of CAD severity （OR： 2.79； 95%CI： 1.72~4.55； P<0.001）. In multivariate logistic regression models， individuals with the lowest quantile of eGDR （T1） were 2.79 times more likely to develop multi-vessel CAD than those with the highest quantile of eGDR （T3） （OR： 2.79； 95%CI： 1.72~4.55； P<0.001）. Multivariate restricted cubic spline analysis showed that eGDR was negatively associated with CAD and multi-vessel CAD （P-nonlinear>0.05）. In non-diabetic patients， compared with the reference group （T3）， the T1 group had a significantly increased risk of CAD （OR： 1.42； 95% CI： 1.00~2.01； P<0.05） and multi-vessel CAD （OR： 1.86； 95%CI： 1.21~2.86； P<0.05）. No statistical association was found between eGDR and CAD in diabetic patients. In ROC curve analysis， when eGDR was added to traditional model for CAD， significant improvements were observed in the model's recognition of CAD and multi-vessel CAD. ConclusionOur study shows eGDR levels are inversely associated with CAD and CAD severity. eGDR， as a non-insulin measure to assess IR， could be a valuable indicator of CAD severity for population.

BACKGROUND:Sepsis-induced systemic inflammation leads to rapid bone mass loss;however,there is a lack of effective treatments.Ulinastatin is an anti-inflammatory drug,but its protective effect and mechanism on bone under sepsis-induced systemic inflammation are still unclear. OBJECTIVE:To explore whether ulinastatin can relieve acute bone loss caused by lipopolysaccharide. METHODS:(1)Animal experiment.Thirty male C57BL/6 mice were randomly divided into three groups(n=10 per group):control group,model group and experimental group.The control group was injected intraperitoneally with normal saline,the model group was injected intraperitoneally with lipopolysaccharide,and the experimental group was injected intraperitoneally with lipopolysaccharide and ulinastatin.In the experimental group,ulinastatin was injected continuously for 3 days.After intraperitoneal injection of ulinastatin for 14 days,femoral tissues were taken for CT scanning and pathological observation.(2)Cell experiment.C57BL/6 mouse primary osteoblasts were isolated and divided into three groups:the control group was routinely cultured,lipopolysaccharide was added to the model group,and lipopolysaccharide with ulinastatin was added to the experimental group.Cell proliferation and osteogenic differentiation were detected.C57BL/6 mouse bone marrow mononuclear cells were isolated and divided into three groups:the control group was routinely cultured,lipopolysaccharide was added to the model group,and lipopolysaccharide and ulinastatin were added to the experimental group.Osteoclast differentiation was detected. RESULTS AND CONCLUSION:(1)Animal experiment.CT scanning and hematoxylin-eosin staining showed that bone mass in lipopolysaccharide-treated mice was reduced but increased after treatment with ulinastatin.Tartrate resistant acid phosphatase staining showed that the number of osteoclasts in bone tissue increased in the model group,but significantly decreased in the experimental group compared with the model group.(2)Cell experiment.Cell counting kit-8 assay showed that lipopolysaccharide treatment inhibited the proliferation of osteoblasts,and ulinastatin elevated the proliferation of osteoblasts after lipopolysaccharide treatment.Alkaline phosphatase staining,alizarin red staining and osteogenesis-related gene(alkaline phosphatase,Runx2,osteocalcin,osteoblastin,nuclear factor κB receptor-activating factor ligand,osteoprotegerin)detection showed that lipopolysaccharide treatment inhibited osteogenic differentiation of osteoblasts and elevated the nuclear factor κB receptor-activating factor ligand/osteoprotegerin ratio;ulinastatin did not have any significant effect on the reduction of osteoblast function induced by lipopolysaccharide but decreased the nuclear factor κB receptor-activating factor ligand/osteoprotegerin ratio.Tartrate resistant acid phosphatase staining and osteoclast-related gene(tartrate resistant acid phosphatase and matrix metalloproteinase 9)detection showed that lipopolysaccharide treatment could promote osteoclast differentiation of bone marrow monocytes,while ulinastatin could inhibit lipopolysaccharide-induced osteoclast differentiation of bone marrow monocytes.(3)Overall,ulinastatin can significantly inhibit lipopolysaccharide-induced bone loss,mainly through promoting osteoblast proliferation and directly or indirectly inhibiting osteoclast differentiation to alleviate bone loss and achieve osteoprotective effects.

Objective:To assess the effectiveness of a 6-month Ba Duan Jin exercise program in improving the balance of community-dwelling older adults.Methods:A two arms, parallel-group, cluster randomized controlled trial was conducted in 1 028 community residents aged 60-80 years in 40 communities in 5 provinces of China. Participants in the intervention group (20 communities, 523 people) received Ba Duan Jin exercise 5 days/week, 1 hour/day for 6 months, and three times of falls prevention health education, and the control group (20 communities, 505 people) received falls prevention health education same as the intervention group. The Berg balance scale (BBS) score was the leading outcome indicator, and the secondary outcome indicators included the length of time of standing on one foot (with eyes open and closed), standing in a tandem stance (with eyes open and closed), the closed circle test, and the timed up to test.Results:A total of 1 028 participants were included in the final analysis, including 731 women (71.11%) and 297 men (28.89%), and the age was (69.87±5.67) years. After the 3-month intervention, compared with the baseline data, the BBS score of the intervention group was significantly higher than the control group by 3.05 (95% CI: 2.23-3.88) points ( P<0.001). After the 6-month intervention, compared with the baseline data, the BBS score of the intervention group was significantly higher than the control group by 4.70 (95% CI: 4.03-5.37) points ( P<0.001). Ba Duan Jin showed significant improvement ( P<0.05) in all secondary outcomes after 6 months of exercise in the intervention group compared with the control group. Conclusions:This study showed that Ba Duan Jin exercise can improve balance in community-dwelling older adults aged 60-80. The longer the exercise time, the better the improvement.

Objective To investigate the effect of DEAD-box RNA helicases(DDX5 helicase)on the proliferation,apoptosis and differentiation of leukemia cell line K562 cells.Methods Data from the Gene Expression Profiling Interactive Analysis(GEPIA)cancer gene database was utilized to analyze the expression of DDX5 mRNA in tis-sues of leukemia patients.Survival curve analysis was conducted to assess the relationship between DDX5 mRNA expression levels and the prognosis of leukemia patients.Small interfering RNA(siRNA)was used for transient transfection of K562 cells to knock down DDX5.Real-time quantitative PCR(RT-PCR)was employed to verify the silencing effect,and Western blot was used to detect protein expression levels.CCK-8 assay was conducted to ex-amine cell proliferation capability,and Western blot and immunofluorescence were utilized to detect the expression levels of cell proliferation-related proteins.Flow cytometry was used to detect cell apoptosis,and the expression lev-els of cell differentiation-related proteins were assessed by flow cytometry and Western blot.Lastly,the effects of DDX5 silencing on the expression levels of P21 and P53 proteins were examined by RT-PCR and Western blot.Re-sults GEPIA database analysis revealed that the expression level of DDX5 in human leukemia bone marrow tissues was significantly higher than that in healthy individuals,and patients with low DDX5 expression had longer survival time.In vitro experiments demonstrated that knocking down DDX5 significantly inhibited the proliferation of K562 cells.Flow cytometry results indicated that DDX5 silencing promoted apoptosis and induced cell differentiation by enhancing the expression of CD11b and CD14 proteins.Furthermore,silencing DDX5 up-regulated the expression levels of P21 and P53 proteins.Conclusion DDX5 may potentially inhibit the proliferation of leukemia cell line K562,promote apoptosis,and induce differentiation by targeting P53.

OBJECTIVE This study aimed to investigate the effects of Buyang Huanwu Decoction(BYHWD)on delaying vascu-lar aging and explore whether the underlying mechanism is associated with microRNA-665(miR-665)/DNA damage-regulated autoph-agy modulator1(DRAM1)-mediated autophagy.METHODS Male SD rats with natural aging were randomly divided into the aging group,BYHWD low,medium,high dosage groups(9.25,18.5,37.0 g·kg-1)and resveratrol group(80 mg·kg-1),with a young group set as well.The rats in each group were dissected and the blood vessels were collected.ELISA was used to assess senescence as-sociated β-galactosidase(SA-β-Gal)activity and advanced glycation end products(AGEs)level in vascular tissues.HE,Masson,and EVG staining were performed to observe the morphological structure of the vascular tissues.The qPCR was performed to detect the expression of miR-665 in vascular tissues.Bioinformatics analysis and dual-luciferase reporter gene experiments were used to validate the targeting relationship between miR-665 and DRAM1.Transmission electron microscope was used to observe the autophagosome.Western blot was performed to determine the protein expression of p16,DRAM1 and autophagy-related proteins Bec-lin1,p62 and LC3.Immunohistochemistry was used to detect the protein expression of DRAM1 in vascular tissues.RESULTS Com-pared to the young group,the aging group showed increased SA-β-Gal activity,AGEs level and p16 protein expression(P<0.01),disordered arrangement of vascular tissues,thickened media,increased collagen fibers,fractured and disorganized elastic fibers.The expression of miR-665 was upregulated(P<0.01).The number of autophagosomes was reduced.The protein expression of Beclin1 and LC3Ⅱ/Ⅰ downregulated(P<0.01),while the protein expression of p62 was upregulated(P<0.01).In addition,the protein ex-pression of DRAM1 was downregulated in vascular tissues(P<0.01).Compared to the aging group,intervention with BYHWD and resveratrol reduced SA-β-Gal activity(P<0.01),AGEs level and p16 protein expression(P<0.05,P<0.01),improved vascular morphology and elastic fiber structure,reduced collagen fibers.High dose BYHWD significantly downregulated miR-665 expression(P<0.01),increased the number of autophagosomes.Different doses of BYHWD significantly upregulated protein expression of Bec-lin1(P<0.05,P<0.01),medium and high doses of BYHWD significantly upregulated protein expression of LC3Ⅱ/Ⅰ(P<0.01),and downregulated protein expression of p62(P<0.01).High dose BYHWD significantly upregulated protein expression of DRAM1 in vascular tissues of aging rats(P<0.05).Bioinformatics analysis revealed the presence of specific complementary binding sites between the sequences of miR-665 and DRAM1.Dual-luciferase reporter assays confirmed that miR-665 targeted DRAM1 gene and negatively regulated DRAM1 protein expression.CONCLUSION BYHWD may promote the protein expression of DRAM1 by inhibiting the ex-pression of miR-665,thereby promoting vascular autophagy and delaying vascular aging.

Objective To explore the prevention and treatment strategies for pancreatic fistula and pancreatic fistula combined with hemorrhage after pancreaticoduodenectomy.Methods We retrospectively reviewed 90 cases of pancreaticoduodenectomy at Guangdong Provincial People's Hospital from August 2019 to December 2022.According to whether postoperative pancreatic fistula occurred,the 90 patients were divided into a postoperative pancreatic fistula group(n=35)and a postoperative non-pancreatic fistula group(n=55).Among the 35 patients with postoperative pancreatic fistula,they were further categorized into two subgroups based on the presence of hemorrhage:the pancreatic fistula with hemorrhage group(n=10)and the pancreatic fistula without hemorrhage group(n=25).Chi-square test or Fisher's exact test was used for univariate analysis.Variables with statistical dif-ferences were selected for stepwise regression variable screening.Multivariate Logistic regression analysis was used to determine the independent risk factors for the occurrence of pancreatic fistula and postoperative pancreatic fistula with hemorrhage.Results All 90 patients successfully completed the pancreaticoduodenectomy.The incidence of postoperative pancreatic fistula was 38.9%(35/90).Significant differences were observed in pancreatic duct diam-eter(P=0.013),intraoperative blood loss(P=0.045),anastomosis type(P=0.045),and residual pancreatic texture(P=0.10)between the two groups(P<0.05).Multivariate logistic regression analysis revealed that soft pancreas texture,pancreatic duct diameter<3 mm,intraoperative blood loss≥300 mL,and pancreaticojejunostomy were independent risk factors for postoperative pancreatic fistula.Among patients with postoperative pancreatic fistula,multivariate logistic regression analysis identified pancreatic fistula volume>100 mL and duration of postop-erative pancreatic fistula>7 days as independent risk factors for hemorrhage.Conclusions The risk of pancreatic fistula after pancreatoduodenectomy is relatively high.Attention to preoperative pancreatic duct diameter and standardized evaluation of pancreatic texture can help identify postoperative pancreatic fistula.Careful hemostasis during operation and avoidance of early postoperative hemorrhage can reduce the incidence of grade B and C pan-creatic fistulas.Patients with pancreatic fistula should be warned of the occurrence of combined hemorrhage when the fistula volume is greater than 100ml and the duration of postoperative pancreatic fistula is greater than 7 days.

Objective:To investigate the molecular characteristics of obacunone, and to screen and identify potential targets of obacunone against sepsis.Methods:The pharmacological parameters and molecular characteristics of obacunone were analyzed with the aid of the Traditional Chinese Medicine Systems Pharmacology Database Analysis Platform (TCMSP). The potential targets of obacunone against sepsis were screened using SwissTargetPrediction and Drug Repositioning and Adverse Drug Reaction Chemical-Protein Interactome (DRAR-CPI) software, with a Z'-score < -0.5. The anti-sepsis targets of obacunone were selected by Online Mendelian Inheritance in Man (OMIM), Comparative Toxicogenomics Database (CTD) and Therapeutic Target Database (TTD). The anti-sepsis potential target was identified by molecular docking software.Results:The oral bioavailability of obacunone was 81.58% and the drug-likeness was 0.57 indicating that obacunone showed good drug formation. A total of 242 potential targets were screened through SwissTargetPrediction and DRAR-CPI software, 13 targets were directly related to sepsis. Cathepsin G (CTSG), caspase-1 (CASP1), S100 calcium binding protein A9 (S100A9), protein C (inactivator of coagulation factors Ⅴa and Ⅷa, PROC), mitogen-activated protein kinase 1 (MAPK1), glucose-6-phosphate dehydrogenase (G6PD), interleukin-10 (IL-10), migration inhibitory factor (MIF), complement C5a receptor 1 (C5AR1), caspase-3 (CASP3), CXC chemokine receptor 2 (CXCR2), thrombin receptor (F2R), nicotinamide phosphoribosyltransferase (NAMPT) were identified as the potential targets for anti-sepsis of obacunone by molecular docking software, the free binding energies were -32.55, 1.26, -30.00, 300.08, -31.88, -30.29, -21.38, -30.79, 16 777.84, -21.80, 6 443.36, -20.38, -23.47 kJ/mol, respectively.Conclusions:Obacunone can inhibit blood coagulation and improve inflammatory response by regulating PROC and F2R. It regulates MIF, S100A9, G6PD and IL-10 to play a role in immune response. It regulates CTSG, CASP1, MAPK1, C5AR1 and CASP3 to protect sepsis-damaged organs. By regulating CXCR2, it can reduce the excessive migration of neutrophils to the site of inflammation, alleviate tissue damage. By regulating NAMPT, it improves cellular energy status, reduces oxidative stress, and protects cells from damage.

The traditional diagnosis of neuropsychiatric disorders mainly depends on the subjective evaluation of specialists,neuropsychological test,biochemical examination and other methods,which lacks objective,accurate and intelligent biomarkers.With the rapid development of neuroimaging and artificial intelligence technology,unsupervised learning has been widely used in the auxiliary diagnosis of neuropsychiatric disorders for it has the advantages of independence of external labels,high model generalization,and automatic feature extraction.Compared with the traditional supervised learning methods,unsupervised learning is more capable of achieving objective,accurate and intelligent diagnosis of neuropsychiatric disorders.Herein an overview on the applications of unsupervised learning in the auxiliary diagnosis of neuropsychiatric disorders is provided,summarizing the findings of unsupervised learning in Alzheimer's disease,schizophrenia,major depressive disorder,and autism spectrum disorder,and discussing the research challenges such as insufficient image processing capability,small sample size,insufficient biochemical index data.The corporation with neural network,multi-site large sample size,and deep fusion of multidimensional data are the development trends of unsupervised learning method.

Purpose: Numerous barriers hinder individuals with mental illness from seeking medical assistance in rural regions, yet a comprehensive understanding of these challenges remains elusive. This metasynthesis aims to understand the barriers and facilitators in medical help-seeking among rural individuals with mental illness. Methods: We systematically searched seven databases [PubMed, CINAHL, Medline (OVID), PsycINFO (OVID), Cochrane, Embase, and ProQuest] in May 2023 and included the studies if they reported the barriers or/and facilitators to seek healthcare in rural patients with mental illness. We conducted hand search and citation search on Google Scholar for literature supplements. Thematic analysis was employed. Results: The study included 27 articles reporting on the barriers and facilitators to seeking medical help in this population from 2007 to 2023. We ultimately identified themes at three levels: navigating the terrain of vulnerability and empowerment (the individual with mental illness), navigating the terrain of external environment (the external environment) and connectivity within the healthcare ecosystem for mental health (the health service system). Conclusions We must design more effective strategies to improve mental healthcare access for rural patients, considering cultural nuances and health service utilization patterns. This requires a multi-level approach, tailored to the unique needs of diverse populations.