Objective To identify the enhancer profile marked by histone H3K27ac modification in high-grade intraepithelial neoplasia(HGIN)in order to reveal the novel regulatory mechanism of HGIN pathogensis.Methods Gastric tissue samples were collected from Department of Gastroenterology of Army Medical Center of PLA between June 2022 and June 2023,including 14 normal gastric tissues(Nor group),31 HGIN tissues(HGIN group)and 17 gastric cancer tissues(GC group).Cleavage under targets and tagmentation(CUT&Tag)technique was employed to capture enhancer regions modified by histone H3K27ac.Multi-omics analysis was performed to identify HGIN-specific active enhancers and their potentially regulated genes.Immunohistochemical profiling was performed to assess differential expression of the gene of interest across clinically stratified specimens,combined with CRISPR-dCas9-mediated ablation of active enhancers to monitor the gene of interest transcriptional dynamics and validate enhancer-mediated regulatory mechanisms.Results Epigenomic sequencing obtained the data with excellent quality,and indicated that obvious remodeling was observed in H3K27ac enhancers in HGIN and GC groups(P<0.05),though no significant difference in the genome-wide distribution of H3K27ac modification among the 3 groups.Combining transcriptome data revealed that enhancer remodeling may up-regulate the expression of the proliferation-related target gene,CD24,in the HGIN tissue;while,inhibiting enhancer activity can notably reduce CD24 expression level(P<0.05).Immunohistochemical assay displayed a positive correlation between the expression levels of CD24 and Ki-67(P<0.001).Conclusion The remodeling of H3K27ac enhancer represents a significant epigenetic feature of the transformation from normal condition to HGIN.Remodeling of H3K27ac enhancer up-regulates CD24,which may facilitate the abnormal proliferation of gastric epithelial cells.

Objective To analyze clinical characteristics of mesenchymal-subtype gastric cancer(Mes-GC)by integrating multi-omics data and explore the characteristics of tumor cells and microenvironment associated with the risk for recurrence.Methods Gastric tumor tissue samples were collected from the patients who visited Department of Gastroenterology of Army Medical Center of PLA from January 2022 to December 2023.Transcriptome and genome sequencing were applied for these tissue samples,including 19 cases of diffuse-type gastric cancer,22 cases of intestinal-type gastric cancer,and 23 cases of mixed-type gastric cancer patients.Bioinformatics analysis was employed to investigate the differences in clinical characteristics and tumor microenvironment between Mes-GC and non-mesenchymal-subtype gastric cancer(non-Mes-GC)by integrating data resources including The Cancer Genome Atlas(TCGA),Gene Expression Omnibus(GEO),and National Genomics Data Center(NGDC).Results Compared to non-Mes-GC patients,Mes-GC ones were characterized by later clinical stages,deeper tumor infiltration,and higher rates of lymph node metastasis.Kaplan-Meier survival analysis confirmed that Mes-GC patients were associated with shorter survival time,poor prognosis as well as increased risk of cancer recurrence(P<0.05).Single-cell RNA sequencing data revealed that tumor cells in Mes-GC showed higher expression levels of the genes related to stemness,metastasis(P<0.05),and epithelial-mesenchymal transition(EMT).And in the tumor microenvironment,there were significant more myeloid cells,smooth muscle cells,endothelial cells and fibroblasts,with the most pronounced elevation in the proportion of fibroblasts(P<0.05).Moreover,the patients with larger proportion of fibroblasts were associated with poorer prognosis.Conclusion Mes-GC tumor cells exhibit higher stemness and EMT characteristics,and stromal cells such as myeloid cells,endothelial cells,and fibroblasts are enriched in the tumor microenvironment.These features may be key factors contributing to poor prognosis and high recurrence rate of Mes-GC.

Objective To identify the enhancer landscape marked by histone H3K27ac modifications in diffuse-type gastric cancer(DGC)tissues,and to elucidate the epigenetic remodeling mechanisms by which active enhancers regulate cachexia-related genes.Methods Gastric mucosal tissue samples were collected from Department of Gastroenterology of Army Medical Center of PLA during January 2022 to March 2023,including 10 normal gastric mucosa tissues(Normal group),10 DGC tissues diagnosed with cachexia(DGC group),and 10 organoids derived from DGC tissues(Organoid group).Using H3K27ac chromatin targeting cleavage and tagmentation(CUT&Tag)technology,genomic modification regions were captured to screen specific active enhancers and their potential target genes in DGC tissues.CRISPR-dCas9 gene editing technology was used to intervene with the enhancers,and the expression of target genes was detected with Western blotting and qRT-PCR.Sixteen female SPF-grade BALB/c Nude mice(6～8 weeks old,weighing 18～21 g)were utilized to establish an orthotopic xenograft tumor model using the human diffuse-type gastric cancer cell line MKN45.Cachexia-related phenotypes were evaluated in 3 groups:normal group(n=4),silencing group(n=6),and control group(n=6).Results Significant differential enhancer regions were identified between DGC and normal gastric mucosa tissues.DGC tissues exhibited a marked increase in enhancer abundance(P<0.05)and signal intensity when compared with the normal counterparts.Integrated analysis of transcriptome data revealed that some of these active enhancers up-regulated the expression of GDF15,a cachexia-associated target gene in DGC.Targeted silencing of the active enhancer of GDF15 using CRISPR/dCas9-KRAB plasmid technology resulted in a significant reduction in GDF15 expression at both mRNA levels(P<0.05)and protein.Results from orthotopic transplantation experiments of DGC demonstrated that silencing of active enhancers alleviated the cachexia phenotype in nude mice(P<0.05).Conclusion DGC exhibits enhancer remodeling,which regulates the expression of the cachexia-associated gene GDF15,and thereby contributes to the pathogenesis and progression of cancer cachexia.

Objective To assess the longitudinal mitral annular plane systolic excursion ( M APSE) of different directions in normal fetuses during mid‐late pregnancy based on two‐dimensional speckle tracking imaging ( ST I) . Methods Seventy‐six normal fetuses during middle and late pregnancy were selected at 26－32 weeks of gestation . T he peak M APSE was measured by free angle M‐mode echocardiography ( FAM ) perpendicular to the lateral annulus in the mitral annular plane . The time‐displacement curves of interventricular septal mitral annulus in three different directions including points A ,B and C through transverse level of apex were recorded by STI . T he peak M APSE of interventricular septal mitral annulus ( SEPT‐M APSE‐A ,SEPT‐M APSE‐B ,SEPT‐M APSE‐C) in three different directions including points A ,B and C and the time to peak ( T T P :SEPT‐T T P‐A ,SEPT‐T T P‐B ,SEPT‐T T P‐C) were recorded respectively . T he time‐displacement curves of lateral mitral annulus in three different directions including points A ,B and C through transverse level of apex were recorded by STI . T he peak M APSE of lateral mitral annulus ( LAT‐M APSE‐A ,LAT‐MAPSE‐B ,LAT‐MAPSE‐C) in three different directions including points A ,B and C ,the time to peak( LA T‐T T P‐A ,LA T‐T T P‐B ,LA T‐T T P‐C) were recorded respectively . Finally ,the data were analyzed statistically . Results T he peak M APSE of the lateral mitral annulus in 3 different directions including points A ,B and C[ LA T‐M APSE‐A ( 3 .62 ± 1 .01) mm ,LA T‐M APSE‐B ( 3 .95 ± 1 .04) mm ,LAT‐M APSE‐C ( 4 .45 ± 1 .05) mm ] were greater than those of the interventricular septum mitral annulus[ SEPT‐MAPSE‐A (3 .41 ± 0 .63)mm ,SEPT‐MAPSE‐B (3 .07 ± 0 .50) mm ,SEPT‐MAPSE‐C (2 .82 ± 0 .51) mm] . LAT‐M APSE‐C and SEPT‐M APSE‐A were the largest longitudinal excursions of mitral annulus . T he differences were statistically significant in points B and C ( P ＜0 .05) . T here was no significant difference in point A ( P ＞0 .05) . LA T‐M APSE‐C was less than FAM‐M APSE [ ( 6 .06 ± 1 .35 ) mm ] . T here was a significant difference between them ( P ＜0 .05 ) . Strong correlation was found between them ( r ＝0 .896 , P＜0 .05) . T here were no significant differences in the time to peak of interventricular septal mitral annulus [ SEPT‐T T P‐A ( 0 .210 ± 0 .008 ) s ,SEPT‐T T P‐B ( 0 .213 ± 0 .008 ) s ,SEPT‐T T P‐C ( 0 .210 ± 0 .005 ) s] in directions including points A ,B ,C ( P＞ 0 .05 ) . T here were no significant differences in time to peak of lateral mitral annulus [ LAT‐T T P‐A ( 0 .210 ± 0 .008 ) s , LAT‐T T P‐B ( 0 .213 ± 0 .006 ) s , LAT‐T T P‐C ( 0 .210 ± 0 .007) s] in directions inclucling points A ,B ,C ( P ＞0 .05) . Conclusions Longitudinal systolic motion of fetal left ventricular wall during mid‐late pregnancy has good synchronization . Longitudinal motion of fetal mitral annulus is a comprehensive movement of multiple directions and different degrees of displacement ,with the movement perpendicular to the annulus as the maximum displacement direction . T he displacement parameters of mitral annulus measured by ST I can reflect the left ventricular longitudinal systolic function and have clinical application value in evaluating the left ventricular longitudinal systolic function of fetuses .

Objective: To assess the longitudinal mitral annular plane systolic excursion (MAPSE) of different directions in normal fetuses during mid-late pregnancy based on two-dimensional speckle tracking imaging (STI). Methods: Seventy-six normal fetuses during middle and late pregnancy were selected at 26-32 weeks of gestation. The peak MAPSE was measured by free angle M-mode echocardiography (FAM) perpendicular to the lateral annulus in the mitral annular plane. The time-displacement curves of interventricular septal mitral annulus in three different directions including points A, B and C through transverse level of apex were recorded by STI. The peak MAPSE of interventricular septal mitral annulus (SEPT-MAPSE-A, SEPT-MAPSE-B, SEPT-MAPSE-C) in three different directions including points A, B and C and the time to peak (TTP: SEPT-TTP-A, SEPT-TTP-B, SEPT-TTP-C) were recorded respectively. The time-displacement curves of lateral mitral annulus in three different directions including points A, B and C through transverse level of apex were recorded by STI. The peak MAPSE of lateral mitral annulus (LAT-MAPSE-A, LAT-MAPSE-B, LAT-MAPSE-C) in three different directions including points A, B and C, the time to peak(LAT-TTP-A, LAT-TTP-B, LAT-TTP-C) were recorded respectively. Finally, the data were analyzed statistically. Results: The peak MAPSE of the lateral mitral annulus in 3 different directions including points A, B and C[LAT-MAPSE-A (3.62±1.01)mm, LAT-MAPSE-B (3.95±1.04)mm, LAT-MAPSE-C (4.45±1.05)mm] were greater than those of the interventricular septum mitral annulus[SEPT-MAPSE-A (3.41±0.63)mm, SEPT-MAPSE-B (3.07±0.50)mm, SEPT-MAPSE-C (2.82±0.51)mm]. LAT-MAPSE-C and SEPT-MAPSE-A were the largest longitudinal excursions of mitral annulus. The differences were statistically significant in points B and C (P<0.05). There was no significant difference in point A (P>0.05). LAT-MAPSE-C was less than FAM-MAPSE[(6.06±1.35)mm]. There was a significant difference between them(P<0.05). Strong correlation was found between them(r=0.896, P<0.05). There were no significant differences in the time to peak of interventricular septal mitral annulus [SEPT-TTP-A (0.210±0.008)s, SEPT-TTP-B (0.213±0.008)s, SEPT-TTP-C (0.210±0.005)s] in directions including points A, B, C(P>0.05). There were no significant differences in time to peak of lateral mitral annulus[LAT-TTP-A(0.210±0.008)s, LAT-TTP-B(0.213±0.006)s, LAT-TTP-C(0.210±0.007)s] in directions inclucling points A, B, C(P>0.05). Conclusions Longitudinal systolic motion of fetal left ventricular wall during mid-late pregnancy has good synchronization. Longitudinal motion of fetal mitral annulus is a comprehensive movement of multiple directions and different degrees of displacement, with the movement perpendicular to the annulus as the maximum displacement direction. The displacement parameters of mitral annulus measured by STI can reflect the left ventricular longitudinal systolic function and have clinical application value in evaluating the left ventricular longitudinal systolic function of fetuses.

Objective To investigate the mechanisms of protective effects of dexmedetomidine on lungs in patients of sepsis complicated with acute respiratory distress syndrome (ARDS). Methods The adult patients with sepsis complicated with ARDS, the oxygenation index (PaO2/FiO2) was 150-200 mmHg (1 mmHg = 0.133 kPa), acute physiology and chronic health evaluationⅡ (APACHEⅡ) score was 10-20, need mechanical ventilation (MV) treatment > 72 hours, and admitted to intensive care unit (ICU) of the First Affiliated Hospital of Guangxi Medical University from September 2013 to June 2017 were enrolled. According to the random number table method, the patients were divided into three groups (n = 80): no sedation group, propofol group (0.3-4.0 mg·kg-1·h-1) and dexmedetomidine group (0.2-0.7 μg·kg-1·h-1). The three groups were adequately analgesic treated with remifentanil. The sedation target was -1-0 of Richmond agitation-sedation score (RASS). The levels of interlenkin-6 (IL-6) and tumor necrosis factor-α (INF-α) were determined by enzyme linked immunosorbent assay (ELISA) before sedation, and 24, 48, 72 hours after sedation. The expressions of inflammatory signaling proteins in bronchoalveolar lavage fluid (BALF) were determined by Western Blot before sedation and 72 hours after sedation. Results There were no significant changes for inflammatory factors in serum, and inflammatory signaling proteins and anti-apoptotic signaling proteins in alveolar exfoliated cells in no sedation group. The levels of IL-6 and TNF-α in serum and the expressions of Toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88) and phosphorylated c-Jun N-terminal kinase (p-JNK) in alveolar cells in propofol group and dexmedetomidine group were all significantly reduced after sedation, moreover, it was more significantly in the dexmedetomidine group compared with propofol group [48 hours: TNF-α (ng/L) was 153.76±29.16 vs. 179.82±30.28;72 hours: IL-6 (ng/L) was 272.18±42.76 vs. 304.49±44.93, TNF-α (ng/L) was 102.18±30.25 vs. 140.28±28.92, TLR4 (IA value) was 0.288±0.034 vs. 0.648±0.029, MyD88 (IA value): 0.356±0.030 vs. 0.752±0.044, p-JNK (IA value): 0.256±0.027 vs. 0.303±0.034, all 1 < 0.05]. The expression of p-Akt in alveolar cells in propofol group and dexmedetomidine group was all significant increased after sedation, moreover, it was more significantly in the dexmedetomidine group compared with propofol group (IA value: 1.032±0.030 vs. 0.743±0.028, 1 < 0.05). Conclusion Dexmedetomidine exerts the protective effects on lungs in patients of sepsis complicated with ARDS through the TLR4-MyD88-JNK signaling pathway.

Objective To explore clinical characteristics of patients with Klinefelter′s syndrome(KS; 47,XXY).Methods 227 male patients with 47,XXY treated by artificial insemination with donor(AID)were included.Age, education, height, weight, body mass index(BMI), testicular volume, FSH, LH, testosterone(T), prolactin(PRL), estradiol(E2)were analyzed retrospectively.Results Of these patients, their height were(176.4±5.5)cm, weight(74.5±12.7)kg, BMI(23.89±3.66)kg/m2[77 of overweight(33.92%)and 34 of obesity(14.98%)], FSH(38.35±14.33)IU/L, LH(19.40±9.00)IU/L, T(132.00±194.50)ng/dl, E2(23.90±15.00)ng/L, PRL(10.50±8.20)μg/L, E2/T 0.21±0.80.Testicular volume had the positive association with the level of T(r=0.197, P=0.003).BMI had the negative association with the serum concentration of T(r=-0.284, P=0.000), while positive association with the E2(r=0.174, P=0.009)and ratio of E2/T(r=0.323, P=0.000).Age had no association with T, E2, and E2/T(P>0.05), but had negative association with the serum concentration of LH(r=-0.154, P=0.02)nd FSH(r=-0.196,P=0.03).The higher education group were older(P<0.01), while the level of T were lower(P<0.01).Conclusion In patients with Klinefelter′s syndrome(KS; 47,XXY), level of T may associate with testicular volume.T, E2, and the ratio of E2/T seem to associate with their height, BMI, and education level.

Objective To study the influence of mild hypothermia on pulmonary vascular permeability in patients with acute respiratory distress syndrome (ARDS) induced by infection.Methods A prospective randomized controlled trial was conducted.Patients with ARDS induced by infection satisfied criteria including age 18-70 years,endotracheal intubation and mechanical ventilation (MV),and without severe coagulation disorder admitted to intensive care unit (ICU) of the First Affiliated Hospital of Guangxi Medical University from May 2012 to November 2015 were enrolled,excluding tumor,burn,cardiac disease,vascular disease,and endovascular surgery within 3 months.The patients enrolled were randomly divided into non-temperature controlled group and mild hypothermia group.The primary diseases in all patients were treated according to the treating principles,including respiratory support,integrated treatment of organ support and symptomatic treatment.Besides,the patients in the mild hypothermia group were administered with systemic hypothermia,and the patients' core body temperature (nasopharyngeal temperature) was rapidly decreased to 34-35 ℃ within 1 hour.Acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score,oxygenation index (PaO2/FiO2),extravascular lung water index (EVLWI) and pulmonary vascular permeability index (PVPI) in two groups at 1,24,48,and 72 hours after treatment or core temperature up to standards were monitored respectively.Enzyme-linked immunosorbent assay (ELISA) was used to determine the levels of vascular endothelial growth factor (VEGF) in venous blood as well as tumor necrosis factor-α (TNF-α) and surfactant apoprotein A (SP-A) in bronchoalveolar lavage fluid (BALF),and circulating endothelial cell (CEC) was counted.The duration of mechanical ventilation and 7-day survival rate were recorded.Results Fifty-six patients were enrolled,with 32 in non-temperature controlled group and 24 in mild hypothermia group.There was no difference in baseline variables including gender,age,APACHE Ⅱ score,PaO2/FiO2 between two groups.APACHE Ⅱ score,EVLWI,PVPI,VEGF,CEC,and TNF-α in both groups were gradually increased with treatment time prolongation,and PaO2/FiO2 and SP-A were gradually decreased.Compared with non-temperature controlled group,APACHE Ⅱ score (16.34±4.27 vs.19.24 ± 5.95),EVLWI (mL/kg:12.17 ± 2.26 vs.12.39 ± 4.71),PVPI (15.40 ± 10.95 vs.16.08 ± 10.24),VEGF (ng/L:127.92 ± 31.49 vs.159.12 ± 40.67),CEC (cells/μL:4.15 ± 1.79 vs.5.70 ± 2.38),and TNF-α (ng/L:147.18 ± 48.85 vs.257.17 ±40.84) in mild hypothermia group were significantly decreased from 24 hours (all P ＜ 0.05),and PaO2/FiO2 [mmHg (1 mmHg =0.133 kPa):175.03± 12.64 vs.162.53 ± 14.15] and SP-A (μg/L:80.85 ± 16.18 vs.62.06 ± 17.28) were significantly increased (both P ＜ 0.05),the duration of mechanical ventilation was significantly shortened (days:10.38 ± 1.50 vs.15.74 ± 3.06,P ＜ 0.01),and 7-day survival rate was significantly increased (75.0％ vs.46.9％,P ＜ 0.05).Conclusion Mild hypothermia can reduce the pulmonary vascular permeability,and improve pulmonary function in early phase in patients with ARDS,as well as shorten the duration of mechanical ventilation,and decrease short-term mortality.

Objective To evaluate regulatory effects of glutamate receptor antagonists on the proliferation and migration of WM451LU malignant melanoma cells, and to explore their related mechanisms. Methods WM451LU cells at exponential growth phase were classified into 3 groups to be treated with the glutamate receptor antagonist MK?801 at 100μmol/L(MK?801 group), the glutamate receptor antagonist CPCCOEt at 10μmol/L(CPCCOEt group), or culture medium(control group). After 24?hour treatment, methyl thiazolyl tetrazolium(MTT)assay was performed to determine cell proliferation rates, scratch assay to evaluate the migration activity of cells, and Western?blot analysis to measure expression levels of proliferating cell nuclear antigen (PCNA), protein kinase Cα(PKCα) both on cell membrane and in cytoplasm, and phosphorylated mitogen?activated protein kinase(p?MAPK). Results After 24?hour treatment, cell proliferation rates were significantly decreased in the MK?801 group and CPCCOEt group compared with the control group(63%± 3.1%and 60%± 2.4%vs. 100%± 1.1%, both P<0.05). The scratch assay showed that cell?free zones in the control group gradually narrowed over time, and the scratch wound tended to close. However, the cell?free zones in the MK?801 group and CPCCOEt group narrowed more slowly compared with the control group, and were still wide after 24?hour culture with no obvious closure of the scratch. The MK?801 group and CPCCOEt group both showed significantly decreased expressions of PCNA(77.0% ± 5.4% and 72.0% ± 4.2% respectively), PKCα on the cell membrane(0.12 ± 0.02 and 0.14 ± 0.02 respectively), and p?MAPK(0.48 ± 0.03 and 0.36 ± 0.04 respectively) compared with the control group(PCNA:100.0%± 1.3%;PKCα:0.38 ± 0.01;p?MAPK:1.00 ± 0.02;all P<0.05).Conclusion In vitro suppression of glutamate receptors can inhibit the proliferation and migration of WM451LU cells, likely through the mediation of the PKCα?MAPK signaling pathway.

Objective To investigate the expression levels of Interleukin(IL)?6 and matrix metalloproteinases(MMP)?13 in synovial fluid by in?vigorating kidney and activating blood formulae in treating rabbit knee osteoarthritis model. Methods A total of 30 New Zealand rabbit were ran?domly divided into blank group,model group,invigorating kidney group,activating blood group and invigorating kidney and activating blood group. Rabbits model with knee osteoarthritis were established by improved Hulth method. To give corresponding respectively the medicinal broth,model group was given saline,knee joint synovial fluid was collected after 4,8 and 12 weeks. Enzyme linked immunosorbent assay(ELISA)was used to measure the levels of IL?6 and MMP?13. Results The levels of IL?6 in rabbit knee osteoarthritis were obviously higher than that of normal control group at both 4 weeks and 8 weeks(P<0.001). But there was no statistical difference on the levels of IL?6 compared with controls in 12 weeks. In addition,the level of MMP?13 at 4 weeks,8 weeks and 12 weeks were significantly higher than the blank control group(P<0.001). After 8 weeks of Chinese medicine administration,the levels of IL?6 in synovial fluid were significantly decreased in invigorating kidney group,activating blood group and invigorating kidney and activating blood group(P<0.001),but there was no statistical difference among groups in 12 weeks. The MMP?13 levels of synovial fluid was significantly lower than the model group(P<0.001). Conclusion Our results indicate that IL?6 and MMP?13 par?ticipate in the pathological development of the rabbit knee osteoarthritis. Invigorating kidney and activating blood formulae could reduce the expres?sion of IL?6 and MMP?13 and alleviate osteoarthritis progression,and which is superior to the pure invigorating kidney formulae and activating blood formulae.