Objective:To investigate the effects of long non-coding RNA 01694(LINC01694)regulating the microRNA-128-3p(miR-128-3p)/telomeric repeat binding factor 1(TERF1)axis on the malignant biological behaviors of prostate cancer(PC)cells.Methods:Cancer tissues and corresponding adjacent tissues from 20 PC patients undergoing surgery at the Department of Urology,Jingzhou Central Hospital,between January 2023 and January 2024 were collected.Human PC cell lines(PC-3,DU145,LNCaP,C4-2)and normal human prostate epithelial RWPE-1 cells were routinely cultured.LNCaP cells were transfected with sh-LINC01694,sh-NC,miR-128-3p inhibitor,inhibitor-NC,miR-128-3pmimics,pcDNA,and pcDNA-LINC01694 using Lipo6000? transfection reagent.Cells were divided into the following groups:Ctrl,sh-NC,sh-LINC01694,sh-LINC01694+NC inhibitor,sh-LINC01694+miR-128-3p inhibitor,pcDNA,and pcDNA LINC01694 groups.The mRNA expression of LINC01694,miR-128-3p,and TERF1 in PC tissues and cells,as well as LNCap cells in each group,was detected by qPCR.The protein expression of TERF1,caspase-3,cyclin D1,E-cadherin,and N-cadherin in LNCaP cells of each group was detected by WB method.Clone formation assay,flow cytometry,and Transwell chamber assay were applied to detect proliferation,apoptosis,migration,and invasion of LNCaP cells,respectively.Dual luciferase reporter gene assay,RNA pull-down assay,and RNA-binding protein immunoprecipitation(RIP)assay were applied to verify the targeting binding relationship between LINC01694 and miR-128-3p as well as between TERF1 and miR-128-3p.Nude mouse LNCaP cell xenograft experiment was conducted to assess the effect of LINC01694 knockdown on tumor growth.Results:LINC01694 was highly expressed in PC tissues and cells(all P<0.05).Knockdown of LINC01694 in LNCaP cells promoted the protein expression of miR-128-3p,caspase-3,and E-cadherin,inhibited the protein expression of LINC01694,TERF1,cyclin D1,and N-cadherin,reduced cell proliferation,migration,and invasion,and promoted apoptosis(all P<0.05).All these effects were partially reversed by the miR-128-3p inhibitor(all P<0.05).LINC01694 could directly bind to miR-128-3p(P<0.05),while miR-128-3p could directly bind to TERF1 mRNA(P<0.05),indicating that LINC01694 regulates the miR-128-3p/TERF1 axis.Knockdown of LINC01694 significantly inhibited the growth of LNCaP cell xenografts in nude mice(P<0.05).Conclusion:LINC01694 regulates the malignant biological behaviors of LNCaP cells through the miR-128-3p/TERF1 axis.

Objective The pathophysiological process of acute high-altitude hypoxia-induced lung injury affects protein expression levels,which are mainly evaluated by Western blot.No systematic study has investigated changes in internal control proteins as calibration loading amounts.Methods Lung injury at an altitude of 6000 m was induced in a low-pressure,low-oxygen chamber for 8,24,and 72 h using C57BL/6J mice.Establishment of the model was confirmed by hematoxylin and eosin staining.Expression levels of various internal control proteins,including vinculin,α-tubulin,eukaryotic translation initiation factor 5(EIF5),β-actin,and glyceraldehyde 3-phosphate dehydrogenase(GAPDH)were detected by Western blot,and total protein expression was detected by Coomassie blue staining.Furthermore,the lung injury model in vitro was establised by using,Bronchial epithelial cell(BZAS-2B)andhunman umbilical vein endothelial cells(HUVECS)confirmed by TUNEL staining.Expression levels of internal control proteins were detected by Western blot,and total protein expression was detected by Coomassie Blue staining.Results Acute 8,24,and 72 h hypoxic models were successfully established in lung tissue,demonstrating consistent total protein expression and stable levels of the internal reference proteins vinculin,α-tubulin,EIF5,andβ-actin.GAPDH expression was elevated in the HH8 h,HH24 h,and HH72 h groups compared with the normoxia(Nor)group,but only the increase at HH72 h groups was significant.Similarly,8,24,and 48 h hypoxic models were successfully established in BEAS-2B cells and HUVECs,with consistent total protein expression.In BEAS-2B cells,expression levels of the internal reference proteins β-actin and GAPDH were consistent with the normoxic control(NC)group,while vinculin,α-tubulin,and EIF5 expression levels were significantly reduced under hypoxic conditions for up to 24 h.In HUVECs,vinculin and α-tubulin expression levels were also consistent with the NC group,while EIF5,β-actin,and GAPDH expression levels were significantly reduced at 8 h and increased at 48 h.Conclusions Acute hypoxia induces lung tissue injury,and protein expression levels of the internal reference proteins vinculin,α-tubulin,EIF5,and β-actin are stable,making them suitable internal references for Western blot.Additionally,Western blot detected differential expression levels of the internal reference proteins vinculin,α-tubulin,EIF5,β-actin,and GAPDH in BEAS-2B cells and HUVECs,as the most important in vitro lung tissue models of hypoxia-induced injury.

Interleukin- (IL) 23 drives pathogenic differentiation of Th17 cells via the JAK2-STAT3 signaling pathway, upregulates IL-17A/IL-22 expression, and disrupts intestinal barrier integrity, thereby playing a pivotal role in the pathogenesis of Crohn's disease (CD). IL-23p19 inhibitors—exemplified by risankizumab, mirikizumab, and guselkumab—precisely block this pathway. Key phase 3 trials have demonstrated their efficacy in CD, showing significant clinical benefits in patients refractory to conventional therapies or biologics, with no new safety signals identified. The ultimate treatment goal for CD is deep healing (mucosal-transmural-biochemical composite remission) as defined by STRIDE II. However, current evidence exhibits critical limitations: absence of head-to-head drug comparisons, insufficient data on biologic-experienced subpopulations, and heterogeneous follow-up durations leading to uncertainties in long-term safety. Future studies require standardized head-to-head trials with enhanced subgroup analyses to optimize precision therapeutics.

Objective The pathophysiological process of acute high-altitude hypoxia-induced lung injury affects protein expression levels,which are mainly evaluated by Western blot.No systematic study has investigated changes in internal control proteins as calibration loading amounts.Methods Lung injury at an altitude of 6000 m was induced in a low-pressure,low-oxygen chamber for 8,24,and 72 h using C57BL/6J mice.Establishment of the model was confirmed by hematoxylin and eosin staining.Expression levels of various internal control proteins,including vinculin,α-tubulin,eukaryotic translation initiation factor 5(EIF5),β-actin,and glyceraldehyde 3-phosphate dehydrogenase(GAPDH)were detected by Western blot,and total protein expression was detected by Coomassie blue staining.Furthermore,the lung injury model in vitro was establised by using,Bronchial epithelial cell(BZAS-2B)andhunman umbilical vein endothelial cells(HUVECS)confirmed by TUNEL staining.Expression levels of internal control proteins were detected by Western blot,and total protein expression was detected by Coomassie Blue staining.Results Acute 8,24,and 72 h hypoxic models were successfully established in lung tissue,demonstrating consistent total protein expression and stable levels of the internal reference proteins vinculin,α-tubulin,EIF5,andβ-actin.GAPDH expression was elevated in the HH8 h,HH24 h,and HH72 h groups compared with the normoxia(Nor)group,but only the increase at HH72 h groups was significant.Similarly,8,24,and 48 h hypoxic models were successfully established in BEAS-2B cells and HUVECs,with consistent total protein expression.In BEAS-2B cells,expression levels of the internal reference proteins β-actin and GAPDH were consistent with the normoxic control(NC)group,while vinculin,α-tubulin,and EIF5 expression levels were significantly reduced under hypoxic conditions for up to 24 h.In HUVECs,vinculin and α-tubulin expression levels were also consistent with the NC group,while EIF5,β-actin,and GAPDH expression levels were significantly reduced at 8 h and increased at 48 h.Conclusions Acute hypoxia induces lung tissue injury,and protein expression levels of the internal reference proteins vinculin,α-tubulin,EIF5,and β-actin are stable,making them suitable internal references for Western blot.Additionally,Western blot detected differential expression levels of the internal reference proteins vinculin,α-tubulin,EIF5,β-actin,and GAPDH in BEAS-2B cells and HUVECs,as the most important in vitro lung tissue models of hypoxia-induced injury.

High mobility group box 1 (HMGB1), when released extracellularly, plays a pivotal role in the development of spinal cord synapses and exacerbates autoimmune diseases within the central nervous system. In experimental autoimmune encephalomyelitis (EAE), a condition that models multiple sclerosis, the levels of extracellular HMGB1 and interleukin-33 (IL-33) have been found to be inversely correlated. However, the mechanism by which IL-33 deficiency enhances HMGB1 release during EAE remains elusive. Our study elucidates a potential signaling pathway whereby the absence of IL-33 leads to increased binding of P300/CBP-associated factor with HMGB1 in the nuclei of astrocytes, upregulating HMGB1 acetylation and promoting its release from astrocyte nuclei in the spinal cord of EAE mice. Conversely, the addition of IL-33 counteracts the TNF-α-induced increase in HMGB1 and acetylated HMGB1 levels in primary astrocytes. These findings underscore the potential of IL-33-associated signaling pathways as a therapeutic target for EAE treatment.
Animals ; Encephalomyelitis, Autoimmune, Experimental/metabolism* ; Astrocytes/metabolism* ; Interleukin-33/metabolism* ; HMGB1 Protein/metabolism* ; Acetylation ; Mice, Knockout ; Mice, Inbred C57BL ; p300-CBP Transcription Factors/metabolism* ; Mice ; Spinal Cord/metabolism* ; Cells, Cultured ; Female ; Signal Transduction

Objective:To observe the clinical efficacy and effects on emotional state, anorectal physiological function, serum inflammation factors and neurotransmitters of repetitive transcranial magnetic stimulation (rTMS) on functional anorectal pain (FAP) patients, and to explore the potential therapeutic mechanisms.Methods:From September 1, 2022 to December 31, 2023, a total of 50 FAP patients who were admitted to Nanjing Hospital of Chinese Medicine Affiliated to Nanjing University of Chinese Medicine were enrolled in this study. The patients were randomly divided into the treatment group (20 cases) and the control group (20 cases) according to a random number table and relevant exclusion criteria. The treatment group received rTMS treatment and the control group received sham rTMS treatment. The Hamilton anxiety scale (HAMA) score, Hamilton depression scale (HAMD) score, visual analogue scale (VAS) score, high-resolution anorectal manometry data (anal resting pressure, anal squeeze pressure, initial sensation shreshold, defecation shreshold, defecation urgency shreshold, and tolerance shreshold), and the levels of serum inflammatory factors (interleukin(IL)-4, IL-8, tumor necrosis factor-α) and 5-hydroxytryptamin(5-HT) were recorded before and after treatment. Independent sample t-test, paired t-test, Mann-Whitney U test and Wilcoxon signed-rank test were used for statistical analysis. Results:The VAS, HAMA, and HAMD scores of the treatment group after treatment were lower than those before treatment (3.85±2.23 vs. 6.85±1.98, 4.40±3.39 vs. 8.75±6.60, and 7.10±6.56 vs. 12.85±7.20), and were also lower than those of the control group after treatment(6.50±1.76, 8.20±6.65, 12.10±6.80), and the differences were statistically significant ( t=5.68, 4.72, 6.06; -4.17, -2.27, -2.37; P<0.001, <0.001, <0.001; <0.001, =0.028, and =0.023). The initial sensation shreshold, defecation shreshold, defecation urgency shreshold, and tolerance shreshold of the treatment group after treatment were higher than those before treatment(30.00(30.00, 46.00) mmHg (1 mmHg=0.133 kPa) vs. 23.00(18.50, 29.00) mmHg, 50.00(44.50, 60.00) mmHg vs. 37.00(30.75, 51.50) mmHg, (74.30±16.02) mmHg vs. (63.70±22.21) mmHg, 119.00(100.00, 148.00) mmHg vs. 98.00 (69.50, 153.00) mmHg), and the tolerance shreshold of the treatment group after treatment was higher than that of the control group after treatment(119.00 (100.00, 148.00) mmHg vs. 102.00(84.50, 111.50) mmHg), and the differences were statistically significant ( Z=–3.14 and –2.86, t=-4.02, Z=-2.84 and -2.11; P=0.002, 0.004, 0.001, 0.004, and 0.035). Additionally, the 5-HT level of the treatment group after treatment was higher than that before treatment (1 549.41 (1 320.21, 1 640.03) μg/L vs. 1 081.52(874.36, 1 626.79) μg/L), and the difference was statistically significant ( Z=-2.88, P=0.004). Conclusion:The rTMS treatment can effectively relieve the pain, anxiety and depression, improve visceral sensitivity, and influence the neurotransmitter level of brain-gut axis in FAP patients.

Cognitive frailty, as one of the independent dimensions of frailty syndrome, has received increasing attention from researchers in recent years. Cognitive frailty not only reduces the quality of life for elderly people, but also increases the risk of adverse outcomes such as falls, disabilities, hospitalization, dementia, and death. This article introduces the concept, assessment methods, influencing factors, and intervention measures of cognitive frailty, emphasizing the important role of general practitioners in screening and management of cognitive frailty.

Cognitive frailty, as one of the independent dimensions of frailty syndrome, has received increasing attention from researchers in recent years. Cognitive frailty not only reduces the quality of life for elderly people, but also increases the risk of adverse outcomes such as falls, disabilities, hospitalization, dementia, and death. This article introduces the concept, assessment methods, influencing factors, and intervention measures of cognitive frailty, emphasizing the important role of general practitioners in screening and management of cognitive frailty.

The diagnosis of hematological disorders is currently established from the combined results of different tests, including those assessing morphology (M), immunophenotype (I), cytogenetics (C), and molecular biology (M) (collectively known as the MICM classification). In this workflow, most of the results are interpreted manually (i.e., by a human, without automation), which is expertise-dependent, labor-intensive, time-consuming, and with inherent interobserver variability. Also, with advances in instruments and technologies, the data is gaining higher dimensionality and throughput, making additional challenges for manual analysis. Recently, artificial intelligence (AI) has emerged as a promising tool in clinical hematology to ensure timely diagnosis, precise risk stratification, and treatment success. In this review, we summarize the current advances, limitations, and challenges of AI models and raise potential strategies for improving their performance in each sector of the MICM pipeline. Finally, we share perspectives, highlight future directions, and call for extensive interdisciplinary cooperation to perfect AI with wise human-level strategies and promote its integration into the clinical workflow.

Interleukin- (IL) 23 drives pathogenic differentiation of Th17 cells via the JAK2-STAT3 signaling pathway, upregulates IL-17A/IL-22 expression, and disrupts intestinal barrier integrity, thereby playing a pivotal role in the pathogenesis of Crohn's disease (CD). IL-23p19 inhibitors—exemplified by risankizumab, mirikizumab, and guselkumab—precisely block this pathway. Key phase 3 trials have demonstrated their efficacy in CD, showing significant clinical benefits in patients refractory to conventional therapies or biologics, with no new safety signals identified. The ultimate treatment goal for CD is deep healing (mucosal-transmural-biochemical composite remission) as defined by STRIDE II. However, current evidence exhibits critical limitations: absence of head-to-head drug comparisons, insufficient data on biologic-experienced subpopulations, and heterogeneous follow-up durations leading to uncertainties in long-term safety. Future studies require standardized head-to-head trials with enhanced subgroup analyses to optimize precision therapeutics.