Objective To map the super-enhancers remodeling of intestinal gastric cancer and reveal the tumor biological functions of the super-enhancers and the downstream target genes that may be activated.Methods A total of 31 normal gastric mucosal tissues,23 intestinal gastric cancer tissues and 9 intestinal gastric cancer organoids were collected from the Department of Gastroenterology of Army Medical Center of PLA from January to December 2022.Chromatin targeting histone H3K27ac modified chromatin targeting cleavage under targets and tagmentation(CUT&Tag)sequencing was conducted on above tissues.The remodeling profiles of super-enhancers in intestinal gastric cancer were analyzed and the key target genes were identified based on bioinformation tools.CRISPRi technology was used to intervene with the super-enhancers,the expression of target genes was detected with Western blotting,and the proliferation,migration and invasion abilities were detected by CCK-8 assay and Transwell chambers in the control group and the intervention group.Results There was a significant difference in the signal of super-enhancers between intestinal gastric cancer tissues and normal gastric mucosal tissues(P<0.05),and the active super-enhancers in cancer tissues may be involved in biological processes such as negative regulation of the immune system and cell adhesion.The expression of up-regulated cell migration-inducing protein(CEMIP)in tumor cells was regulated by the super-enhancers,and intervening the super-enhancers down-regulated the expression of CEMIP(P<0.05),and inhibited the cell proliferation,invasion and migration abilities of tumor cells(P<0.05).Conclusion Super-enhancer remodeling is observed in intestinal gastric cancer,and they can up-regulate the expression of CEMIP gene and promote the growth,migration and invasion of cancer cells.

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.

BACKGROUND:Carnosic acid,a bioactive compound found in rosemary,has been shown to reduce inflammation and reactive oxygen species(ROS).However,its mechanism of action in osteoclast differentiation remains unclear. OBJECTIVE:To investigate the effects of carnosic acid on osteoclast activation,ROS production,and mitochondrial function. METHODS:Primary bone marrow-derived macrophages from mice were extracted and cultured in vitro.Different concentrations of carnosic acid(0,10,15,20,25 and 30 μmol/L)were tested for their effects on bone marrow-derived macrophage proliferation and toxicity using the cell counting kit-8 cell viability assay to determine a safe concentration.Bone marrow-derived macrophages were cultured in graded concentrations and induced by receptor activator of nuclear factor-κB ligand for osteoclast differentiation for 5-7 days.The effects of carnosic acid on osteoclast differentiation and function were then observed through tartrate-resistant acid phosphatase staining,F-actin staining,H2DCFDA probe and mitochondrial ROS,and Mito-Tracker fluorescence detection.Western blot and RT-PCR assays were subsequently conducted to examine the effects of carnosic acid on the upstream and downstream proteins of the receptor activator of nuclear factor-κB ligand-induced MAPK signaling pathway. RESULTS AND CONCLUSION:Tartrate-resistant acid phosphatase staining and F-actin staining showed that carnosic acid dose-dependently inhibited in vitro osteoclast differentiation and actin ring formation in the cell cytoskeleton,with the highest inhibitory effect observed in the high concentration group(30 μmol/L).Carnosic acid exhibited the most significant inhibitory effect during the early stages(days 1-3)of osteoclast differentiation compared to other intervention periods.Fluorescence imaging using the H2DCFDA probe,mitochondrial ROS,and Mito-Tracker demonstrated that carnosic acid inhibited cellular and mitochondrial ROS production while reducing mitochondrial membrane potential,thereby influencing mitochondrial function.The results of western blot and RT-PCR revealed that carnosic acid could suppress the expression of NFATc1,CTSK,MMP9,and C-fos proteins associated with osteoclast differentiation,and downregulate the expression of NFATc1,Atp6vod2,ACP5,CTSK,and C-fos genes related to osteoclast differentiation.Furthermore,carnosic acid enhanced the expression of antioxidant enzyme proteins and reduced the generation of ROS during the process of osteoclast differentiation.Overall,carnosic acid exerts its inhibitory effects on osteoclast differentiation by inhibiting the phosphorylation modification of the P38/ERK/JNK protein and activating the MAPK signaling pathway in bone marrow-derived macrophages.

Objective:To investigate the protective mechanism of physical exercise in regulating the transition of microgli-al phenotype and cognitive impairment in Alzheimer's disease(AD).Method:Male 5xFAD mice at the age of 5 months were randomly divided into physical exercise(PE)and control group,and 5-month-old male C57BL/6J mice were taken as wild type group.Three months of wheel-running exercise was conducted in PE group.After completion of exercise program,the anxiety-like behavior and cognitive impairment were assessed by Morris water maze test and open field test.The protein expression of α/β-hydrolase domain-containing 12(ABHD12)was detected by Western Blots.The cellular location of AB-HD12 was detected by immunofluorescence staining.In vitro study,BV-2 cells were transfected with lentivirus overexpression or knockdown ABHD12.Aβ-42 oligomer was incubated to establish the AD model in vitro.RT-qPCR、western blots and immunofluorescence staining were performed to verify the up-and down-regulation of ABHD12,as well as to explore the effect of ABHD12 in the transition of microglial phenotype.Result:Physical exercise ameliorated the cognitive and emotional dysfunction in 5xFAD mice.Physical exer-cise also reduced Aβ deposition and maintained the synaptic plasticity.Mechanically,physical exercise up-regu-lated the expression of ABHD12,which is expressed in microglia,but not in astrocytes or neurons.Physical exercise also increased the number of M2 microglia and decreased the number of M1 microglia.The knock-down of ABHD12 exacerbates the inflammatory response of Aβ oligomer induced BV-2 cells.Conclusion:Physical exercise ameliorated cognitive dysfunction in Alzheimer's disease by up-regulating AB-HD12,prompting the transition of microglial phenotype to inhibit the inflammation,and to reduce the Aβ de-position.

Objective:To investigate the protective mechanism of physical exercise in regulating the transition of microgli-al phenotype and cognitive impairment in Alzheimer's disease(AD).Method:Male 5xFAD mice at the age of 5 months were randomly divided into physical exercise(PE)and control group,and 5-month-old male C57BL/6J mice were taken as wild type group.Three months of wheel-running exercise was conducted in PE group.After completion of exercise program,the anxiety-like behavior and cognitive impairment were assessed by Morris water maze test and open field test.The protein expression of α/β-hydrolase domain-containing 12(ABHD12)was detected by Western Blots.The cellular location of AB-HD12 was detected by immunofluorescence staining.In vitro study,BV-2 cells were transfected with lentivirus overexpression or knockdown ABHD12.Aβ-42 oligomer was incubated to establish the AD model in vitro.RT-qPCR、western blots and immunofluorescence staining were performed to verify the up-and down-regulation of ABHD12,as well as to explore the effect of ABHD12 in the transition of microglial phenotype.Result:Physical exercise ameliorated the cognitive and emotional dysfunction in 5xFAD mice.Physical exer-cise also reduced Aβ deposition and maintained the synaptic plasticity.Mechanically,physical exercise up-regu-lated the expression of ABHD12,which is expressed in microglia,but not in astrocytes or neurons.Physical exercise also increased the number of M2 microglia and decreased the number of M1 microglia.The knock-down of ABHD12 exacerbates the inflammatory response of Aβ oligomer induced BV-2 cells.Conclusion:Physical exercise ameliorated cognitive dysfunction in Alzheimer's disease by up-regulating AB-HD12,prompting the transition of microglial phenotype to inhibit the inflammation,and to reduce the Aβ de-position.

Objective To evaluate clinical efficacy of lung transplantation for lung chronic graft-versus-host disease (cGVHD) after hematopoietic stem cell transplantation (HSCT). Methods Clinical data of 12 patients undergoing lung transplantation for lung cGVHD were retrospectively analyzed. Preoperative clinical manifestations and involved organs of patients were analyzed. The lung function before and after lung transplantation was compared, and the survival of patients after lung transplantation was analyzed. Results Eleven patients underwent HSCT due to primary hematological malignancies, including 9 cases of leukemia, 1 case of myelodysplastic syndrome, 1 case of lymphoma. And 1 case underwent HSCT for systemic lupus erythematosus. Among 12 cGVHD patients, skin involvement was found in 8 cases, oral cavity involvement in 5 cases, gastrointestinal tract involvement in 4 cases and liver involvement in 3 cases. All 12 patients developed severe respiratory failure caused by cGVHD before lung transplantation, including 9 cases of typeⅡ respiratory failure and 3 cases of type Ⅰ respiratory failure. Two patients underwent right lung transplantation, 2 cases of left lung transplantation and 8 cases of bilateral lung transplantation. The interval from HSCT to lung transplantation was 75 (19-187) months. Upon the date of submission, postoperative follow-up time was 18 (7-74) months. Ten patients survived, 1 died from severe hepatitis at postoperative 22 months, and 1 died from gastrointestinal bleeding at postoperative 6 months. No recurrence of primary diseases was reported in surviving patients. Conclusions Lung transplantation is an efficacious treatment for lung cGVHD after HSCT, which may prolong the survival time and improve the quality of life of the recipients.

Objective To explore the clinical features and treatment of thyroid carcinoma in children. Method The clinical data of 19 children under 14 years old with thyroid carcinoma diagnosed and treated from January 2003 to January 2014 were retrospectively analyzed. Results In 19 cases (12 males and 7 females), there were 18 cases of papillocarcinoma and one case pf follicular carcinoma. Unilateral lobectomy plus isthmectomy was performed in 6 cases, subtotal thyroidectomy in 4 cases and total thyroidectomy in 9 cases. Unilateral cervical lymph node dissection was performed in 5 cases and bilateral in 11 cases. After the operation, multiple lesions were confirmed by pathology in 9 cases, thyroid capsular invasion in 14 cases, lymphatic metastasis in 15 cases and distant metastasis in 5 cases. All the patients were treated with TSH, and 10 cases were treated with 131I after operation. The median follow-up time was 63 months. There was no death in all cases, while local residual tumor recurrence was found in 2 cases and cervical lymph node metastasis in 2 cases and distant metastasis in one case. Conclusion Thyroid carcinoma in children is mostly well-differentiated, so the overall prognosis is better. However, children who have extracapsular invasion, multiple lesions in bilateral thyroid, cervical lymph node metastasis and distant metastasis are at high risks and should be treated with comprehensive therapy that includes total thyroidectomy.

OBJECTIVE:To develop a method for the determination of valsartan concentration in human plasma and urine. METHODS:Plasma sample were acidified and extracted with diethyl ether for analysis,and urine sample was diluted directly for analysis. The samples were all determined by LC-MS/MS,and the separation was performed on a Aglient ZORBAX SB-C18 column with mobile phase consisted of acetonitrile and 0.1% formic acid (gradient elution) at flow rate of 0.2 ml/min. Ion transition was determined ESI ion source under multiple ion reaction monitoring with quantitative pair m/z 436.4→253.2 and qualitative ion pair m/z 436.4→291.3 for valsartan,and quantitative pair m/z 423.4→207.1 and m/z 423.4→180.2 for internal standard losartan. RE-SULTS:The linear range of valsartan were 4-5 000 ng/ml in plasma and 20-50 000 ng/ml in urine;the limit of quantification were 4 ng/ml and 20 ng/ml;plasma extraction recovery of valsartan were 61.21%-70.30%. The variation coefficient of internal standard normalized matrix effect were 3.20% and 11.21%. The within-day and between-day RSDs were no more than 8.34%. CONCLU-SIONS:The method is proved to be rapid and sensitive,and suitable for the determination of valsartan in human plasma and urine and pharmacokinetics study.

Objective To explore the effects of the diet safety instructions and interventions on aspiration pneumonia in elderly patients with dysphagia. Methods The multidisciplinary team was established involved clinical physicians, dietitians, therapists and nurses. The study investigated 40 elderly long-stay patients with dysphagia who were capable of oral feeding were selected according with the inclusive criteria. After the swallowing assessment and diet safety instructions, the participants were divided into intervention group (n=20) and control group (n=20) under the principle of voluntariness and matching selection. The patients of intervention groups were supplied with all mushy diet; the patients of control group had semi-solid, thick liquid, partial mushy diet, etc. depended on participants′ swallowing situations and tastes. Both groups of patients were oral feeding on their own or with help. The incidence of bucking, food refusal and feeding time per meal were investigated during the three months, and the aspiration pneumonia incidence of two groups were retrospectively analyzed three months before interventions. Results In the intervention group, the incidence of bucking (109), food refusal (22) and feeding time per meal [(18. 64 ± 5. 05) min] were lower than those in the control group [bucking (272);food refusal (135); feeding time per meal (25. 78 ± 8. 37) min] (χ2 =7. 369, 4. 433;t= -4. 363;P<0. 05). After three months′interventions, the incidence of aspiration pneumonia in both groups was decreased that the intervention group was 13 ∶ 4 and the control group was 11 ∶ 7 (χ2 =20. 742,13. 809;P<0. 05). The control group had two cases with bucking aspiration and one cases with multiple organ failure and death after nasal feeding; the intervention group had no dropout. Conclusions A close cooperation of multi-discipline team can significantly improve the detection rate and feeding safety management of elderly patients with dysphagia. The diet intervention and safety eating management can reduce the incidence of aspiration pneumonia and promote the patients′quality of life.

Objective To investigate the pharmacokinetics of cephapirin sodium in healthy volunteers.Methods Twelve healthy volunteers were enrolled and ad ministered with single doses of 0.5,1.0,4.0 g or multiple doses of 1.0 g cephapirin sodium injection by intravenous drip infusion.The concentrations of cephapirin in human plasma and urine were determined by HPLC-MS/MS.The main pharmacokinetic parameters were calculated with WinNonLin 6.3 software. Results The main pharmacokinetic parameters of cephapirin after single dose of 0.5,1.0,4. 0 g and 1.0 g multiple dose cephapirin sodium injection were as follows:Cmaxwere (34.86 ±6.93),(74.77 ±24.23),(319.0 ±44.5),(89.26 ±28.04)μg/mL,AUC0-twere (12.86 ±3.46),(28.31 ±7.46),(163.21 ±34.57),(27.30 ±7.22)μg/(mL·h),t1/2were (0.55 ±0.21),(0.72 ±0.22),(0.71 ±0.27), (0.72 ±0.25),accumulative urine excretion rate of 8 h(1 g)was (44.9 ±12.66)%.Conclusion The process of cephapirin in the dosage range of 0.5 ~4.0 g show linear dynamic feature.There is no accumulation after multiple dosing.Cephapirin sodium was much eli minated from urine in parent drug.