OBJECTIVES: To study the molecular mechanisms of LDH-loaded si-NEAT1 for regulating paclitaxel resistance and tumor-associated macrophage (TAM) polarization in breast cancer. METHODS: qRT-PCR and Western blotting were used to detect the expression of lncRNA NEAT1, miR-133b, and PD-L1 in breast cancer SKBR3 cells and paclitaxel-resistant SKBR3 cells (SKBR3-PR). The effects of transfection with si-NEAT1 and miR-133b mimics on MRP, MCRP and PD-L1 expressions and cell proliferation, migration and apoptosis were investigated using qRT-PCR, Western blotting, scratch and Transwell assays, and flow cytometry. Rescue experiments were conducted using si-NEAT1 and miR-133b inhibitor. Human THP-1 macrophages were cultured in the presence of conditioned media (CM) derived from SKBR3 and SKBR3-PR cells with or with si-NEAT1 transfection for comparison of IL-4-induced macrophage polarization by detecting the surface markers. LDH@si-NEAT1 nanocarriers were constructed, and their effects on MRP, MCRP and PD-L1 expressions and cell behaviors of the tumor cells were examined. THP-1 cells were treated with the CM from LDH@si-NEAT1-treated tumor cells, and the changes in their polarization were assessed. RESULTS: SKBR3-PR cells showered significantly upregulated NEAT1 and PD-L1 expressions and lowered miR-133b expression as compared with their parental cells. Transfection with si-NEAT1 and miR-133b mimics inhibited viability, promoted apoptosis and enhanced MRP and BCRP expressions in SKBR3-PR cells. NEAT1 knockdown obvious upregulated miR-133b and downregulated PD-L1, MRP and BCRP expressions. The CM from SKBR3-PR cells obviously promoted M2 polarization of THP-1 macrophages, which was significantly inhibited by CM from si-NEAT1-transfected cells. Treatment with LDH@si-NEAT1 effectively inhibited migration and invasion, promoted apoptosis, and reduced MRP, BCRP and PD-L1 expressions in the tumor cells. The CM from LDH@si-NEAT1-treated SKBR3-PR cells significantly downregulated Arg-1, CD163, IL-10, and PD-L1 and upregulated miR-133b expression in THP-1 macrophages. CONCLUSIONS LDH@si-NEAT1 reduces paclitaxel resistance of breast cancer cells and inhibits TAM polarization by targeting the miR-133b/PD-L1 axis.
Humans ; MicroRNAs/genetics* ; RNA, Long Noncoding/genetics* ; Paclitaxel/pharmacology* ; Breast Neoplasms/metabolism* ; Drug Resistance, Neoplasm ; B7-H1 Antigen/metabolism* ; Cell Line, Tumor ; Female ; Tumor-Associated Macrophages ; Apoptosis ; Cell Proliferation ; Macrophages ; Cell Movement

Five new flavan-4-ol glycosides jixueqiosides A-E (1-5) and two new flavan glycosides jixueqiosides F and G (6 and 7), along with twelve known flavan-4-ol glycosides (8-19), were isolated from the roots of Pronephrium penangianum. Comprehensive spectral analyses, X-ray single-crystal diffraction, and theoretical electronic circular dichroism (ECD) calculations established structures and absolute configurations. A single crystal structure of flavan-4-ol glycoside (14) was reported for the first time, while the characteristic ECD and NMR data for all isolated flavan-4-ol glycosides (1-5 , 8-19) were analyzed, establishing a set of empirical rules. Activity screening of these isolates showed that 8 and 9 could inhibit the proliferation of MDA-MB-231 and MCF-7 cells with IC₅₀ values of 7.93 ? 2.85 ?mol?L^-1 and 5.87 ? 1.58 ?mol?L^-1 (MDA-MB-231), and 2.21 ? 1.38 ?mol?L^-1 and 3.52 ? 1.55 ?mol?L^-1 (MCF-7), respectively. Western blotting and flow cytometry analyses demonstrated that 8 and 9 dose-dependently induced apoptosis in MDA-MB-231 cells by up-regulating BAX, activating caspase-3 and down-regulating BCL-2. Additionally, compound 8 affected autophagy-related proteins, increasing the ratio of LC3-II/LC3-I and Beclin-1 levels to inhibit MDA-MB-231 cell proliferation. Moreover, anti-inflammatory studies indicated that 2, 3, 7, 13, 14, and 18 moderately inhibited tumor necrosis factor-a (TNF-a), interleukin-6 (IL-6), and nitric oxide (NO) release.
Humans ; Plant Roots/chemistry* ; Glycosides/isolation & purification* ; Anti-Inflammatory Agents/isolation & purification* ; Flavonoids/isolation & purification* ; Cell Proliferation/drug effects* ; Antineoplastic Agents, Phytogenic/isolation & purification* ; Molecular Structure ; Apoptosis/drug effects* ; Cell Line, Tumor ; Tumor Necrosis Factor-alpha/immunology* ; Drugs, Chinese Herbal/pharmacology* ; Interleukin-6/immunology* ; Animals ; Mice

Objective To explore the mediating role of mindful attention and awareness in depressive symptoms and insomnia severity among patients with comorbid depression and insomnia.Methods A cross-sectional study was conducted,enrolling 267 patients with comorbid depression and insomnia who were treated in the outpatient Department of Medical Psychology of Second Affiliated Hospital of Army Medical University,from March to May 2024.Basic demographic and clinical data were collected using a general information questionnaire.Depressive symptom severity was measured via the Patient Health Questionnaire-9(PHQ-9),insomnia severity via the Insomnia Severity Index(ISI),perceived stress via the Perceived Stress Scale-10(PSS-10),and mindful attention and awareness via the Mindful Attention Awareness Scale(MAAS).Pearson correlation analysis was used to examine the correlations between depressive severity,insomnia severity,perceived stress,and mindful attention and awareness.Mediation analysis was performed using Process 4.1.Results The PHQ-9 score was(13.80±5.98)and the ISI score was(17.10±5.56)in the 267 patients.Pearson correlation analysis showed that depressive severity and insomnia severity were positively correlated with perceived stress(r=0.531,0.351,P<0.001)and negatively correlated with mindful attention and awareness(r=-0.373,-0.350,P<0.001).Mediation analysis using Process 4.1 indicated that the combined mediating effect of mindful attention and awareness and insomnia between perceived stress level and depressive level was 0.157,with a 95%confidence interval(CI)of 0.102～0.217,and the total mediating effect was significant(P<0.001).Conclusion Perceived stress directly positively affects depression and indirectly exacerbates depression through insomnia as a mediator,and mindful attention and awareness can weaken the promoting effect of perceived stress on insomnia.

Objective: To investigate the therapeutic effect of bone marrow mesenchymal stem cells ( BMSCs) mod- ified by bone morphogenetic protein ( BMP) -2 gene combined with nano-hydroxyapatite ( nHA) / polyamide 66 ( PA66) on femoral nonunion in rats． Methods : Rat BMSCs were isolated and divided into the stent + BMSCs group，stent + BMSCs + rhBMP-2 group，and stent + BMSCs + Ad-BMP-2 group; human recombinant BMP-2 ( rh- BMP-2) or adenovirus-infected BMP-2 ( Ad-BMP-2) vector was transfected into BMSCs and loaded onto nHA / PA66 stent materials．Flow cytometry was used to detect that BMSCs expressed specific surface markers．Cell pro- liferation was detected by MTT assay，related bioactive factors［platelet-derived growth factor ( PDGF) ，transfor- ming growth factor-β ( TGF-β) ，vascular endothelial growth factor ( VEGF) ，fibroblast growth factor ( FGF) ，os- teocalcin ( OCN) ，osteonectin ( ON) ］were detected by ELISA，alkaline phosphatase ( ALP) activity was detec- ted，and cell growth and adhesion were observed by scanning electron microscopy ( SEM) ．In the atrophic nonun- ion model of SD rats，the stent + BMSCs + rhBMP-2 or stent + BMSCs + Ad-BMP-2 complex was implanted into the defect area，which was divided into the rhBMP-2 group and the Ad-BMP-2 group，and the therapeutic effect was e- valuated by X-ray and MicroCT． Results: The surface of the nHA / PA66 stent was smooth and porous，and BMSCs adhered well．Flow cytometry showed high expression of CD29 and CD90 and low expression of CD45 and CD34 in BMSCs．MTT showed that the cells proliferated rapidly after 72 h．Compared with the stent + BMSCs group and the stent + BMSCs + rhBMP-2 group，the ALP activity，the expressions of PDGF，TGF-β , VEGF，FGF，OCN，and ON in the stent + BMSCs + Ad-BMP-2 group were significantly up-regulated ( P＜0. 05) ．12 weeks after surgery， the stent complex in the Ad-BMP-2 group of rats was completely wrapped by newly formed cortical bone，and the surface was smooth and intact．X-ray showed that the complex in the Ad-BMP-2 group was completely wrapped by newly formed cortical bone，and the recovery degree was better than that in the rhBMP-2 group．Micro-CT results showed that compared with the rhBMP-2 group，the bone volume fraction，trabecular thickness，trabecular num- ber，bone mineral density，and bone mineral content in the Ad-BMP-2 group all increased 12 weeks after surgery ( P＜0. 05) ，and the trabecular separation level decreased ( P＜0. 05) ． Conclusion Ad-BMP-2-transfected BM- SCs combined with nHA / PA66 stent material can express bioactivity more effectively，provide a continuous and good microenvironment for the proliferation and differentiation of BMSCs，which is beneficial to promoting local os- teogenic activity and bone defect repair．

Objective To establish an orthotopic transplantation tumor model of pancreatic cancer derived from transgenic LSL-KrasG12D/+ LSL-Trp53R172H/+ Pdx1-Cre(KPC)mice.To provide a stable and reliable drug preclinical research animal model to study the developmental mechanism and treatment strategies of pancreatic cancer.Methods Tumor tissue derived from KPC transgenic mice with spontaneous pancreatic cancer was transplanted into the C57BL/6J mouse pancreas.Ultrasound was used to monitor tumor growth.HE and immunofluorescence staining was used to evaluate the pathological characteristics of this model.Results The tumor derived from KPC mice grew steadily on the pancreas of C57BL/6J mice.Tumor cell proliferation index Ki67,matrix fibrosis marker αSMA,and immune cell markers CD45 and CD206 were all stably expressed in the tumor.The model stably retained the pathological features of primary pancreatic cancer.Widespread tumor metastases,which were similar to those observed in patients with pancreatic cancer,developed in this model.Conclusions An orthotopic transplantation model derived from a transgenic mouse with spontaneous pancreatic cancer was established successfully.The model simulates the stromal environment and immune cell infiltration of pancreatic cancer and retains strong stability and uniformity with the original tumor.It can be used as an effective drug preclinical research model to study pancreatic cancer progression and treatment strategies.

Pheochromocytomas and paragangliomas(PPGLs)cause symptoms by altering the circulation levels of catecholamines and peptide hormones.Currently,the diagnosis of PPGLs relies on diagnostic imaging and the detection of catecholamines.In this study,we used ultra-performance liquid chromatography(UPLC)/quadrupole time-of-flight mass spectrometry(Q-TOF MS)analysis to identify and measure the perioperative differential metabolites in the plasma of adrenal pheochromocytoma patients.We identified differentially expressed genes by comparing the transcriptomic data of pheochromocytoma with the normal adrenal medulla.Through conducting two steps of metabolomics analysis,we identified 111 differential metabolites between the healthy group and the patient group,among which 53 metabolites were validated.By integrating the information of differential metabolites and differentially expressed genes,we inferred that the cysteine-methionine,pyrimidine,and tyrosine metabolism pathways were the three main metabolic pathways altered by the neoplasm.The analysis of transcription levels revealed that the tyrosine and cysteine-methionine metabolism pathways were downregulated in pheochromocytoma,whereas the pyrimidine pathway showed no significant difference.Finally,we developed an optimized diagnostic model of two metabolites,L-dihydroorotic acid and vanylglycol.Our results for these metabolites suggest that they may serve as potential clinical biomarkers and can be used to supplement and improve the diagnosis of pheochromocytoma.

Objective To analyze the expression of CREM in gastric cancer(GC)and its correlation with prognosis of the patients.Methods TCGA and GEO databases were used to analyze the expression levels of CREM mRNA in GC and adjacent tissues.Immunohistochemistry was used to examine the expression of CREM protein in 43 pairs of GC and adjacent tissues,and the correlation of CREM expression with clinicopathological features of the patients was analyzed.Kaplan-Meier survival analysis was used to explore the relationship between CREM expression and survival of GC patients.LinkedOmics database was used to annotate the GO function and KEGG pathway enrichment of CREM-related genes.Results Database analysis showed that CREM was highly expressed in GC tissues(P<0.05)and positively correlated with poor prognosis in GC patients(P=0.01).Immunohistochemistry results showed significantly higher CREM expression in GC tissues than in paired adjacent tissues(P<0.0001),and its expression level was correlated with T-stage and N-stage of the tumor(P<0.05).The overall survival of GC patients with high expression of CREM was shorter(RR=4.02,P=0.0046).Gene enrichment analysis showed that high CREM expression promotes occurrence and progression of GC very likely through the cell adhesion signaling pathway.Conclusion CREM is highly expressed in GC,and its high expression is associated with a poor prognosis of GC patients,suggesting the potential of CREM to serve as a prognostic indicator for GC.

Objective To analyze the expression of CREM in gastric cancer(GC)and its correlation with prognosis of the patients.Methods TCGA and GEO databases were used to analyze the expression levels of CREM mRNA in GC and adjacent tissues.Immunohistochemistry was used to examine the expression of CREM protein in 43 pairs of GC and adjacent tissues,and the correlation of CREM expression with clinicopathological features of the patients was analyzed.Kaplan-Meier survival analysis was used to explore the relationship between CREM expression and survival of GC patients.LinkedOmics database was used to annotate the GO function and KEGG pathway enrichment of CREM-related genes.Results Database analysis showed that CREM was highly expressed in GC tissues(P<0.05)and positively correlated with poor prognosis in GC patients(P=0.01).Immunohistochemistry results showed significantly higher CREM expression in GC tissues than in paired adjacent tissues(P<0.0001),and its expression level was correlated with T-stage and N-stage of the tumor(P<0.05).The overall survival of GC patients with high expression of CREM was shorter(RR=4.02,P=0.0046).Gene enrichment analysis showed that high CREM expression promotes occurrence and progression of GC very likely through the cell adhesion signaling pathway.Conclusion CREM is highly expressed in GC,and its high expression is associated with a poor prognosis of GC patients,suggesting the potential of CREM to serve as a prognostic indicator for GC.

Objective:To explore the effect of multidisciplinary management combined with 60-second high-risk diabetic foot screening in diabetic foot.Methods:From January to December 2022, 138 patients with diabetic foot were selected from Xinxiang Central Hospital by convenience sampling. The patients were randomly divided into a control group and an observation group, with 69 cases in each group. Control group implemented routine follow-up management of diabetic foot, and observation group carried out multidisciplinary management combined with 60-second high-risk diabetic foot screening on the basis of control group, and the intervention lasted for six months. The progress of Wagner grading of diabetic foot and foot self-care were compared between the two groups.Results:After intervention, the number of Wagner grading progression patients in observation group and control group was four cases (5.80%) and 10 cases (14.49%), respectively. The number of progression patients in observation group was less than that in control group, and the difference was statistically significant (χ 2=4.161, P=0.041). The total score and dimension scores of diabetic foot self-management in the two groups after the intervention were higher than those before the intervention, but only the scores of observation group before and after the intervention were statistically significant ( P<0.05). After intervention, the total score and dimension scores of diabetic foot self-management in observation group were higher than those in control group, with a statistically significant difference ( P<0.05) . Conclusions:Multidisciplinary management combined with 60-second high-risk diabetic foot screening can effectively delay the progress of diabetic foot and improve patients' foot self-care.

Purpose To explore the ability of proton density fat fraction(PDFF)and decay constant T2* values in MRI to reflect skeletal muscle aging.Materials and Methods 3T MRI data of skeletal muscle in the middle thigh of 211 healthy adults from the Third Affiliated Hospital of Southern Medical University from August to December 2023 were prospectively collected.Gender,age,height,weight and body mass index(BMI)were recorded.PDFF value and T2* value of thigh skeletal muscle were measured at post-processing workstation,and statistical differences among different age,gender and BMI groups were analyzed.The correlation between PDFF value and T2* value of thigh skeletal muscle and age and BMI was analyzed.Results There were statistically significant differences in PDFF values of thigh skeletal muscle among different age groups(H=18.476-85.619,all P<0.01).There were significantly differences in T2*values of the left and right quadriceps muscles,hamstrings and adductors among different age groups(H=13.342-47.566,all P<0.05).There were statistically significant differences in the PDFF values of right quadriceps,left and right hamstring,adductor and sartor muscles between male and female groups(Z=-4.929--1.626,all P<0.05),while there were statistically significant differences in T2* values of left sartor muscle(Z=-2.971,P=0.003).There was no statistical significance in PDFF value of skeletal muscle of thigh in different BMI groups(P>0.05),but there were statistically significant differences in T2* value of left and right quadriceps muscle,hamstring muscle and adductor muscle(H=9.542-24.495,all P<0.05).There was a moderate positive correlation between age and PDFF value of thigh skeletal muscle(r=0.635,P<0.01),but a slight negative correlation with T2* value of left and right quadriceps,hamstring and sarcoleus(r=-0.451--0.189,all P<0.01).There was a slight positive correlation between BMI and T2* values of thigh skeletal muscle(r=0.317,P<0.01).There was a moderate negative correlation between the PDFF value and T2* value of all thigh skeletal muscles(r=-0.749--0.624,P<0.01).The PDFF and T2* values of the front and back thigh muscles(quadriceps,hamstring)were most significantly correlated with age and BMI.Conclusion PDFF based on MRI can reflect the age-related changes in the microenvironment of thigh skeletal muscle,and is a potential imaging biological marker for accurate and non-invasive quantitative evaluation of thigh skeletal muscle aging.