Objective To draw the genome-wide distribution and remodeling characteristics of H3K27me3 silencers in signet-ring cell carcinoma of the stomach(SRCC)through epigenetic sequencing technology,and to investigate their roles in transcriptional regulation in order to elucidate the regulatory mechanism of SRCC malignant progression.Methods The study was conducted on 35 gastric samples obtained by gastroendoscopic biopsy(15 normal and 20 SRCC tissues)from Department of Gastroenterology of Army Medical Center of PLA between January 2021 and December 2023.Multi-omics analyses,including assay for transposase-accessible chromatin with high-throughput sequencing(ATAC-seq),cleavage under targets and tagmentation(CUT&Tag)and transcriptome sequencing(RNA-seq),were performed to identify chromatin accessibility,H3K27me3 silencer regions,and transcriptional changes,with aid of Illumina NovaSeq 6000.H3K27me3 related differentially expressed genes(|Log2FC|>1,FDR<0.05)were screened using DESeq2.Gene Ontology(GO)analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis were employed to analyze the enrichment function,and Homer was employed to identify transcription factor motifs.A regulatory network was constructed using Cytoscape,and then validated using immunohistochemistry to explore its regulatory mechanism.Results H3K27me3 silencers were primarily located in distal intergenic regions(37.06%)in SRCC.Compared with the normal tissues,SRCC showed a significant reduction in H3K27me3 silencer signals(95%CI:1.34～2.30,P=0.007)with 6 257 lost sites(FDR<0.01).Integrating CUT&Tag and RNA-seq revealed 380 up-regulated immune-related genes,particularly in T cell receptor signaling(OR=4.2,95%CI:2.8～6.3,P=0.002).Immunohistochemistry confirmed elevated expression of transcription factor EHF(P<0.05).Conclusion There is the remodeling of H3K27me3 silencers in SRCC,and EHF may potentially play a crucial role in the SRCC malignant progression.

Objective:To establish an HPLC method for the determination of N,N'-diacetyl-L-cystine in com-pound amino acid injection.Methods:The HPLC method parameters were as follows,The Atlantis dC18 column(4.6 mm × 150 mm,3 μm),The mobile phase was aammonium formate solution(315 mg ammonium formate was taken and dissolved with 960 mL water)-acetonitrile-methanoic acid(970∶30∶1)as a mobile phase,at a flow rate of 0.7 mL·min-1,and the detection wavelength of 210 nm.Results:The linear range of N,N'-diacetyl-L-cystine was 2.697-53.94 μg·mL-1(r=0.999 9),the limits of detection and quantification were 1.4 μg·mL-1 and 4.4 μg·mL-1 respectively.The average recovery was 100.2％with RSD of 0.5％.Conclusion:The method was proved to be suitable for the determination of N,N'()-diacetyl-L-cystine in compound amino acid injection.

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease affecting both upper and lower motor neurons in the brain and spinal cord. One important aspect of ALS pathogenesis is superoxide dismutase 1 (SOD1) mutant-mediated mitochondrial toxicity, leading to apoptosis in neurons. This study aimed to evaluate the neural protective synergistic effects of ginsenosides Rg1 (G-Rg1) and conditioned medium (CM) on a mutational SOD1 cell model, and to explore the underlying mechanisms. We found that the contents of nerve growth factor, glial cell line-derived neurotrophic factor, and brain-derived neurotrophic factor significantly increased in CM after human umbilical cord mesenchymal stem cells (hUCMSCs) were exposed to neuron differentiation reagents for seven days. CM or G-Rg1 decreased the apoptotic rate of SOD1^G93A-NSC34 cells to a certain extent, but their combination brought about the least apoptosis, compared with CM or G-Rg1 alone. Further research showed that the anti-apoptotic protein Bcl-2 was upregulated in all the treatment groups. Proteins associated with mitochondrial apoptotic pathways, such as Bax, caspase 9 (Cas-9), and cytochrome c (Cyt c), were downregulated. Furthermore, CM or G-Rg1 also inhibited the activation of the nuclear factor-kappa B (NF-κB) signaling pathway by reducing the phosphorylation of p65 and IκBα. CM/G-Rg1 or their combination also reduced the apoptotic rate induced by betulinic acid (BetA), an agonist of the NF-κB signaling pathway. In summary, the combination of CM and G-Rg1 effectively reduced the apoptosis of SOD1^G93A-NSC34 cells through suppressing the NF-κB/Bcl-2 signaling pathway (Fig. 1 is a graphical representation of the abstract).
Humans ; NF-kappa B/metabolism* ; Ginsenosides/pharmacology* ; Amyotrophic Lateral Sclerosis/genetics* ; Culture Media, Conditioned/pharmacology* ; Superoxide Dismutase-1 ; Neurodegenerative Diseases ; Neurons/metabolism* ; Apoptosis

Objective To explore the association between diabetes mellitus type Ⅱ (DM) and deglutition function of amyotrophic lateral sclerosis (ALS) patients.Methods Seventy-five ALS patients older than 45 years and admitted to our hospital from August 2008 to November 2015 were selected into this study;67 of them were without DM and 8 were with DM.Water swallow test,amyotrophic lateral sclerosis severity scale-swallow (s-ALSSS),amyotrophic lateral sclerosis functional rating scale (ALSFR-R) and videofluoroscopic swallow study (VFS) were performed to evaluate the deglutition functions of these ALS patients with or without DM,and the results of the two groups were compared.Results (1) Patients with DM had significantly higher s-ALSSS scores,ingurgitation part in ALSFRS-R scores and in parts of VFS scores,such as transportation to pharyngeal,pharyngeal transit,flow into pharyngeal before reflexion,epiglottic vallecula residue,and piriform sinus residue than patients without DM (8.88±1.34 vs.7.54±1.47,3.50±0.54 vs.2.96±0.77,2.88±0.35 vs.2.16±0.69,2.75±0.46 vs.2.09±0.69,2.88±0.35 vs.2.42±0.56,2.88±0.35 vs.2.39±0.58,P＜0.05).(2) Scores of Kubota drinking test,ALSFRS-R,VGF (including oral phase,pharyngeal phase and aspiration degree) in patients without DM were 2.15±1.12,7.18±1.41(1.78±0.69,1.69±0.60 and 3.72±0.65),and those in patients with DM were 1.88±1.34,8.00±0.93(2.13±0.64,2.00±0.53 and 4.00±0.00);no significant differences were noted between the two groups (P＞0.05),but there was a trend showing that DM patients had higher scores in these evaluations.Conclusion As compared with ALS patients without DM,ALS patients with DM get more mildly impaired deglutition function.

Objective To explore the clinical and electrophysiological features and pulmonary function of 8 Chinese patients with Kennedy's disease (KD) and to enhance the understanding for the disease.Methods Eight patients with KD,admitted to out hospital from July 2010 to December 2013 and confirmed by gene examination,were chosen in our study; their clinical and electrophysiological features,and laboratory characteristics and pulmonary function were compared with those in the healthy volunteers.Results The average age of onset in the 8 patients was (45.13±17.47) years,and the average age of diagnosis was (55.63±12.11) years.The most common complaint was leg weakness.All patients presented hemifacial spasm,tongue muscle atrophy and fibrillation,amyotrophy and gynaecomastia; creatine kinase level was elevated and endocrine disorder appeared in different degrees.The electromyogram characteristics included widespread neurogenic changes accompanied with/without sensory or motor conduction abnormalities.The forced vital capacity,forced expiratory volume in first second,maximal voluntary ventilation and peak expiratory flow rate were significantly lower,and the residual volume in the KD patients was statistically higher than that in the healthy volunteers (P＜0.05).Respiratory muscle strength (maximum inspiratory pressure and maximum expiratory pressure) decreased in the KD patients.Conclusion KD is a degenerative disease with slow clinical progression which has its own characteristics of inheritance pattern and natural course; the age of onset,repeat number of CAG sequences,pulmonary function and respiratory muscle strength may be valuable for illness evaluation.