ObjectiveTo investigate the epigenetic mechanism of Diwu Yanggan Capsule in improving liver regeneration microenvironment in a rat model of liver cancer by regulating DNA methylation， and to provide a basis for scientific clinical medication. MethodsA total of 48 specific pathogen-free Sprague-Dawley rats were divided into normal group， model group， and Diwu Yanggan Capsule group using a random number table， with 16 rats in each group. The Solt-Farber two-step method was used to establish a rat model of liver cancer. The rats in the Diwu Yanggan Capsule group were given Diwu Yanggan Capsule at a dose of 750 mg/kg/d by gavage， and those in the normal group and the model group were given an equal volume of normal saline by gavage. Liver tissue samples were collected from each group of rats after 16 weeks of continuous intervention； DNA methylation chips were used to analyze the change in DNA methylation in liver tissue， and gene ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） analyses were used for data analysis. In addition， the MeDIP-PCR technique was used to detect the changes in candidate differentially methylated genes such as YWHAB， ADCK2， ERLIN2， SEMA3B， and TPH2 in the liver tissue of rats， and Western blot and RT-qPCR were used to verify the expression of key methylated genes. The independent-samples t test was used for comparison of continuous data between two groups， and a one-way analysis of variance was used for comparison between multiple groups， while the least significant difference t-test was used for further comparison between two groups. ResultsThe DNA methylation chip analysis showed that compared with the normal group， the model group had significant methylation changes in the promoter region of 2 422 genes in liver tissue of rats. The GO functional enrichment analysis and the KEGG pathway enrichment analysis showed that these differentially methylated genes were significantly enriched in metabolic pathways such as steroid hormone biosynthesis and drug metabolism-cytochrome P450. Compared with the model group， the Diwu Yanggan Capsule group had significant reversal of promoter methylation in 1 650 genes， and the KEGG enrichment analysis showed that these genes were mainly involved in the pathways closely associated with cell proliferation， apoptosis， and microenvironment regulation， such as the calcium ion signaling pathway， the cAMP signaling pathway， and the extracellular factor signaling pathway. Compared with the model group， the Diwu Yanggan Capsule group had a significant increase in the promoter methylation level of the ADCK2 gene （P<0.05） and significant reductions in the promoter methylation levels of the ERLIN2 and TPH2 genes （all P<0.05）. Compared with the model group， the Diwu Yanggan Capsule group had significant reductions in the mRNA expression levels and the protein expression levels of the ADCK2 （all P<0.05）. ConclusionAbnormal DNA methylation in liver tissue participates in the development and progression of liver cancer. The effect of Diwu Yanggan Capsule on DNA methylation level is an important epigenetic mechanism for its effect in the prevention and treatment of liver cancer.

Objective:To investigate the neuropsychological and imaging features of patients with posterior cortical atrophy (PCA).Methods:Patients of PCA, dementia with Lewy bodies (DLB), typical Alzheimer′s disease (t-AD) who were diagnosed in the Department of Neurology, Huashan Hospital, Fudan University from September 27, 2019, to September 24, 2021 were enrolled, and the normal controls who visited the Outpatient and Physical Examination Centers of Huashan Hospital, Fudan University and Rizhao People′s Hospital at the same time were enrolled, too. Neuropsychological assessments, magnetic resonance imaging (MRI), and positron emission tomography (PET)/CT data of the 4-group subjects were collected. Variance analysis was used to compare the differences in neuropsychological performance among the 4 groups, and the imaging features of PCA patients were summarized.Results:Eleven PCA patients, 17 DLB patients, 31 t-AD patients, and 11 normal controls were included in the study. The cognitive function of patients in the PCA group [Mini-Mental State Examination (MMSE) score 13.52±1.81; Montreal Cognitive Assessment (MoCA) score 7.06±1.72] was significantly impaired compared to the normal control group (MMSE score 27.85±1.75, t=-6.561, P<0.001; MoCA score 23.60±1.59, t=-7.968, P<0.001]. However, there was no statistically significant difference compared to the DLB group and the t-AD group. Patients in the PCA group exhibited more severe impairments in attention, executive function, and language compared to the DLB group (Trail Making Test A score: 298.86±16.16 vs 110.07±18.62, t=9.980, P<0.001; Trail Making Test B score: 305.51±18.89 vs 230.34±23.59, t=2.865, P=0.024; Boston Naming Test score: 8.67±1.53 vs 15.66±1.56, t=-2.682, P=0.013) and the t-AD group (148.91±12.77, t=7.071, P<0.001; 200.78±19.34, t=3.789, P=0.004; 15.15±1.05, t=-2.544, P=0.016). Scores for visuospatial function [PCA group: 1(0, 1), normal control group: 3(3, 3), Z=-4.023, P<0.001] and visual perception [PCA group: 0(0, 1), normal control group: 35(34, 36), Z=-3.704, P<0.001] were significantly lower in the PCA group compared to the normal control group. The cranial MRI findings of PCA patients showed atrophy of the parietal and occipital lobes, with less obvious atrophy of the medial temporal lobe, which can be distinguished from t-AD. 18F-fluorodeoxyglucose PET/CT of the PCA patients showed a relative reduced glucose metabolism in the bilateral parietal lobe, occipital lobe and posterior cingulate gyrus, while the 18F-florbetapir PET/CT showed deposition of amyloid protein in the bilateral frontal lobe, parietal lobe, temporal lobe, and cingulate gyrus. Conclusions:PCA patients exhibit neuropsychological characteristics of visuospatial dysfunction, along with impairments in various cognitive domains such as memory, attention, and executive functions. The typical MRI feature is parietal occipital lobe atrophy, and the PET/CT findings are consistent with metabolic changes in AD.

Considering that photodynamic therapy (PDT)-induced oxygen consumption and microvascular damage could exacerbate hypoxia to drive more glycolysis and angiogenesis, a novel approach to potentiate PDT and overcome the resistances of hypoxia is avidly needed. Herein, morpholine-modified PEGylated bilirubin was proposed to co-deliver chlorin e6, a photosensitizer, and diclofenac (Dc). In acidic milieu, the presence of morpholine could enable the nanocarriers to selectively accumulate in tumor cells, while PDT-generated reactive oxidative species (ROS) resulted in the collapse of bilirubin nanoparticles and rapid release of Dc. Combining with Dc showed a higher rate of apoptosis over PDT alone and simultaneously triggered a domino effect, including blocking the activity and expression of lactate dehydrogenase A (LDHA), interfering with lactate secretion, suppressing the activation of various angiogenic factors and thus obviating hypoxia-induced resistance-glycolysis and angiogenesis. In addition, inhibition of hypoxia-inducible factor-1α (HIF-1α) by Dc alleviated hypoxia-induced resistance. This study offered a sequentially responsive platform to achieve sufficient tumor enrichment, on-demand drug release and superior anti-tumor outcomes in vitro and in vivo.

Objective To investigate the expression and clinical significance of miR-126 and vascular endothelial growth factor (VEGF) in proliferative diabetic retinopathy (PDR).Methods 226 cases of diabetic retinopathy (DR) patients admitted in our hospital were studied,including 110 cases of PDR (group PDR),116 non proliferative diabetic retinopathy (NPDR) (group NPDR).80 patients with diabetes mellitus without retinopathy (NDR) were enrolled in DR group at the same period,another 80 healthy subjects (control group) were selected as control group.The plasma miR-126 level of all subjects was detected by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR).The enzyme linked immunosorbent assay (ELISA) method was used to detect plasma VEGF level.The clinical diagnostic value of miR-126 and VEGF in PDR patients was further analyzed.Results Plasma levels of miR-126 in PDR group,NPDR group and NDR group were lower than those in control group (P ＜ 0.05),PDR group was lower than NPDR group and NDR group (P ＜ 0.05);plasma levels of VEGF in PDR group,NPDR group and NDR group were higher than those in control group (P ＜ 0.05),PDR group was higher than NPDR group and NDR group (P ＜ 0.05).Total cholesterol (TC),triglyceride (TG) and glycated hemoglobin (HbAlc) in PDR group were higher than those in NPDR group and NDR group (P ＜0.05),low density lipoprotein cholesterol (LDL-C) in PDR group,NPDR group and NDR group were higher than those in control group (P ＜ 0.05),and LDL-C in PDR group was higher than that in NPDR group and NDR group (P ＜0.05).High density lipoprotein cholesterol (HDL-C) in PDR group and NDR group was lower than that in control group (P ＜ 0.05),C-reactive protein (CRP) in PDR group,NPDR group and NDR group was higher than that in control group (P ＜ 0.05);plasma miR-126 levels in PDR group were negatively correlated with TC,LDL-C,CRP and HbA1c (P ＜ 0.05),but positively correlated with HDL-C (P ＜ 0.05),and no correlation with TG;the plasma levels of VEGF in PDR patients were positively correlated with TC,TG,LDL-C,CRP and HbAlc (P ＜0.05),but negatively correlated with the expressions of miR126 (r =-0.573,P =0.000);the AUC of miR-126 was 0.861,and when the cut-off value was ＜0.64,the diagnostic sensitivity and specificity were 82.50％,83.64％;the area under curve (AUC) of VEGF was 0.889,and when the cut-off value was ＜ 7.000,the sensitivity and specificity of diagnosis were 82.73％,86.25％;0.847 for the AUC of HbA1c,and the sensitivity and specificity 81.82％,87.50％ respectively.Conclusions Plasma miR-126 is low-expressed and VEGF is high-expressed in PDR patients,there is a negative correlation between the two indexes.They may be involved in the course of PDR through abnormal lipid metabolism and inflammatory reaction and may be a potential biomarker for early diagnosis of PDR.

Objective To explore the effect of behavior training on the rehabilitation of homeless patients with schizophrenia .Methods Totals of 72 homeless patients with schizophrenia were randomly divided into study group and control group with 36 cases in each group .Both groups received the routine treatment and care, and the study group received behavior training in addition .The Nurse Observation Scale for Inpatient Evaluation(NOSIE)and the Quality of Life Questionnaire for Psychiatric patients (QOL-P)were used to evaluate the rehabilitation effectiveness before and 8 weeks after behavior training .Results There were no differences in NOSIE and QOL-P scores in patients before behavior training ( P >0.05 ).While 8 weeks after behavior training, NOSIE scores of personal hygiene (19.06 ±3.15),social interest (16.89 ±2.64) and social competence (20.11 ±2.79)in study group were higher than(14.44 ±2.71),(12.28 ±2.71),(13.67 ±2.22) in control group,and the differences were statistically significant (t=6.656,7.314,10.086,respectively;P<0.01).Irritation(12.00 ±3.28)and retardation(8.61 ±2.78)scores were significantly lower than (17.33 ± 2.99),(14.00 ±2.53)in control group(t=7.218,8.602,respectively;P<0.01).The total score of QOL-P (54.58 ±8.77) in study group was lower than (77.78 ±5.49) in control group,and the difference was statistically significant(t=13.443,P<0.01).Conclusions Behavior training helps to improve the life ability and social function of homeless patients with schizophrenia .It has a positive effect for enhancing the quality of life of patients and promoting the rehabilitation of the disease.

OBJECTIVETo examine whether calcineurin/NFAT signaling pathway mediates endothelin-1 (ET-1)-induced proliferation of pulmonary artery smooth muscle cells (PASMCs) by regulating phosphodiesterase-5 (PDE5) and the effect of the selective calcineurin inhibitor cyclosporine A and PDE5 inhibitor sildenafil on ET-1-induced PASMC proliferation.

METHODSPASMCs were treated with ET-1 to stimulate their proliferation with or without prior treatment of the cells with CsA or sildenafil. Calcineurin activity in the cells was measured using a calcineurin activity assay kit, PDE5 expression examined using immunoblotting, and cGMP level detected using a cGMP direct immunoassay kit. PASMC proliferation following the treatments was determined using [(3)H]thymidine incorporation assay.

RESULTSET-1 caused a 2.05-fold increase in the cellular calcineurin activity, a 1.80-fold increase in PDE5 expression, and a 3.20-fold increase in the DNA synthesis rate, and reduced the cGMP level by 67%. Pretreatment of the cells with Cyclosporine blocked the effects of ET-1, and PDE5 inhibition by sildenafil pretreatment also abolished ET-1-induced reduction of cGMP level in the cells. Both Cyclosporine and sildenafil suppressed ET-1-stimulated PASMC proliferation.

CONCLUSIONActivation of calcineurin/NFAT signaling pathway mediates ET-1-induced PASMC proliferation by stimulating PDE5 expression, which further degrades cGMP. Both Cyclosporine and sildenafil can suppress ET-1-stimulated PASMC proliferation in vitro.

Animals ; Calcineurin ; metabolism ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cyclic GMP ; metabolism ; Cyclic Nucleotide Phosphodiesterases, Type 5 ; metabolism ; Cyclosporine ; DNA ; biosynthesis ; Endothelin-1 ; pharmacology ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; cytology ; enzymology ; NFATC Transcription Factors ; metabolism ; Piperazines ; Pulmonary Artery ; cytology ; Purines ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Sildenafil Citrate ; Sulfones

Pressure sores result from many risk factors, which influence each other. In this paper, various aspects of the development of pressure sores in recent years were summarized, consisted of the main risk factors, pathogenesis, treatment and care methods. To control pressure sores should focus on prevention, and reflect the integrity and the target population.

Objective To isolate the rat insulin gene enhancer binding protein 1(Islet-1)gene and construct plEGFP-C1-Islet-1 recombinant retroviral expression vector.Methods The cDNA encoding the rat Islet-1 gene was isolated by RT-PCR method,the cDNA was first cloned into PGET-1 TA vector to facilitate the sequence and then subcloned into the retroviral vector plEGFP-C1.plEGFP-C1-Islet-1 was transfected into PA317 packaging cells with lipofectamine 2000.Transformants were selected in medium containing G418.Results A 1 050 bp DNA fragment was obtained by RT-PCR;plEGFP-C1-Islet-1 recombinant retroviral expression vector was identified by restrictive enzymes digestion,PA317 cells transfected with recombinant vector expressed enhancer green fluorescent protein(EGFP).Conclusion The gene encoding the rat Islet-1 is obtained and plEGFP-C1-Islet-1 expression vector is constructed successfully.

Objective To study the condition of pheochromocytoma cells(PC12 cells)differentiation into neurons induced by nerve growth factor(NGF).Methods There were five groups :10,20,50,and 80 ?g?L-1 NGF groups and control group which contained 10% fetal calf serum.PC12 cells were exposed to NGF with different concentrations for 72 h and the lengh of neurite and max diameter of PC12 cells were observed at 24,48,and 72 h,respectively.The expression of MAP2 was detected by immunocytochemistry after PC12 cells were exposed to NGF for 72 h.Results The number of MAP2 positive cells was significantly increased in 20,50,and 80 ?g?L-1 groups compared with control group and the most significant concentration was 50 ?g?L-1(P

Objective To study the condition of pheochromocytoma cells(PC12 cells) differentiated into neuron-like cells induced by retinoic acid(RA) and to observe the lengh of neurite and max diameter and the expression of MAP2 in PC12 cells during their differentiation into neuron-like cells.Methods There were seven groups :0.1,0.3,0.5,1.0,2.0 and 5.0 mg?L-1RA groups and control group which contained 10% fetal calf serum.PC12 cells were exposed to RA with different concentrations for 72 h and the lengh of neurite and max diameter of PC12 cells were observed at 24,48,and 72 h respectively.The expression of MAP2 was detected by immunocytochemistry after PC12 cells were exposed to RA for 72 h.Results The number of MAP2 positive cells was significantly increased in 0.3,0.5,1.0,2.0 mg?L-1RA groups compared with control group and the most significant concentration was 1.0 mg?L-1(P