Objective To explore the genome-wide distribution of histone H3K27ac in intestinal type gastric cancer,analyze remodeling features of enhancers and regulome and construct a prediction model for prognosis.Methods H3K27ac CUT&Tag sequencing and RNA sequencing were performed in intestinal type gastric cancer tissues from 15 patients and normal gastric mucosa tissues from 18 healthy volunteers.Bioinformatics analysis was performed to identify the differences in genome distribution of H3K27ac modifications.Based on the distribution characteristics of H3K27ac,the enhancer elements were identified and the remodeling characteristics of enhancer and related regulome were explored.The prediction model for prognosis based on enhancer related target genes was constructed by univariate Cox and multivariate Cox regression analyses.Results The histone H3K27ac modification was mainly distributed in the enhancer region and displayed no significant differences in the genomic distribution patterns between normal and cancer tissues.Compared with normal gastric mucosa,the level of enhancer H3K27ac modification was higher in intestinal type gastric cancer.A total of 8847 enhancers with increased activity in intestinal type gastric cancer were identified,accounting for 8.3%of all enhancers,which might promote malignant behaviors such as proliferation and adhesion of gastric cancer cells.A prognosis-predicting model established based on a panel of 6 genes that upregulated by the acquired enhancer in cancers,which was able to predict the overall survival of patients.Conclusion Enhancer remodeling is one of the significant epigenetic features of intestinal type gastric cancer.These enhancers may drive malignant growth and adhesion of cancer cells by upregulating the expression of MYC,E2F3 and other genes.A prognosis model based on enhancer target genes is constructed.

Objective·To analyze the expression of mitochondrial related gene SFXN3(sideroflexin 3)in head and neck squamous cell carcinoma(HNSCC)and its effect on cell proliferation.Methods·Mitochondrial related genes and TCGA-HNSCC dataset were obtained from public databases to identify the target gene SFXN3 by using bioinformatic methods.The expression of SFXN3 in HNSCC patient samples was analysed by using the UALCAN database,and survival analyses of patients with different SFXN3 expression levels were performed according to TCGA-HNSCC cohort and GEO cohorts(GSE65858,GSE41613 and GSE27020).By using TCGA-HNSCC cohort and GEO cohorts(GSE40020,GSE210287),the differences in SFXN3 expression levels between therapeutic responders and non-responders were compared.Quantitative real-time PCR(qRT-PCR)was used to verify the expression of SFXN3 in the collected HNSCC tumor and para-tumor tissues,and the expression level of SFXN3 in human normal oral epithelial cells and HNSCC tumor cell lines was detected.With the construction of stable-SFXN3-knockdown HNSCC cell lines,the effect of SFXN3 on the proliferation ability of HNSCC cells was observed by using the Incucyte live-cell imaging analysis system and plate colony formation assay.Transcriptome sequencing was performed on the total RNA of stably-SFXN3-knockdown cells and control cells,and pathway enrichment analysis was performed on the genes with differentially down-regulated expression in knockdown group compared with control group.Results·SFXN3 was highly expressed in tumor tissues of HNSCC patients(P=0.000),and the prognosis of patients in the SFXN3-high-expression group was poor(all P＜0.05).The expression level of SFXN3 in the non-responders was higher than that in the responders(P=0.008),indicating an unfavorable prognosis.High expression of SFXN3 was verified in the collected HNSCC tumor tissues and HNSCC cell lines(all P＜0.05).After the knockdown of SFXN3 expression,the proliferation ability of HNSCC cells decreased,and the number of plate clones decreased(all P＜0.05).RNA sequencing showed that the differentially expressed down-regulated genes in HNSCC cells in the SFXN3-knockdown group were enriched in DNA replication,cell cycle,mitochondrial translation,mitochondrial RNA metabolic process and mitochondrial gene expression pathways.Conclusion·SFXN3 is highly expressed in HNSCC and it has negative correlation with the prognosis of patients.The proliferation ability and plate colony formation ability are inhibited in SFXN3-knockdown HNSCC cells,and these may work by affecting mitochondria function.

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

Oolong tea is a semi-fermented tea with strong flavor, which is widely favored by consumers because of its floral and fruity aroma as well as fresh and mellow taste. During the processing of oolong tea, withering is the first indispensable process for improving flavor formation. However, the molecular mechanism that affects the flavor formation of oolong tea during withering remains unclear. Transcriptome sequencing was used to analyze the difference among the fresh leaves, indoor-withered leaves and solar-withered leaves of oolong tea. A total of 10 793 differentially expressed genes were identified from the three samples. KEGG enrichment analysis showed that the differentially expressed genes were mainly involved in flavonoid synthesis, terpenoid synthesis, plant hormone signal transduction and spliceosome pathways. Subsequently, twelve differentially expressed genes and four differential splicing genes were identified from the four enrichment pathways for fluorescence quantitative PCR analysis. The results showed that the expression patterns of the selected genes during withering were consistent with the results in the transcriptome datasets. Further analysis revealed that the transcriptional inhibition of flavonoid biosynthesis-related genes, the transcriptional enhancement of terpenoid biosynthesis-related genes, as well as the jasmonic acid signal transduction and the alternative splicing mechanism jointly contributed to the flavor formation of high floral and fruity aroma and low bitterness in solar-withered leaves. The results may facilitate better understanding the molecular mechanisms of solar-withering treatment in flavor formation of oolong tea.
Camellia sinensis/genetics* ; Gene Expression Profiling ; Plant Leaves ; Plant Proteins/metabolism* ; Taste ; Tea ; Transcriptome/genetics*

Resin cements have been widely employed for bonding all-ceramic restorations in clinical practice, its color stability is directly related to long-term prosthetic effect of restorations. Discoloration of resin cements can be attributed to two causes: endogenous factors are generally related to material compositions and initiation mechanism of polymerization; exogenous factors are mainly related to stimulation of local oral environment. Color stability of resin cements has close relationship with esthetic effect of all-ceramic restorations. The aim of this literature review was to make a presentation and discussion systematically about color stability of resin cements commonly used clinically, its influence factors and influence on all-ceramic restorations, so as to provide a reference for the application of all-ceramic restorations.

Objective To observe humoral and cellular immune responses induced in mice by a TSOL18 recombinant Mycobacterium smegmatis (MS) vaccine of Taenia solium.Methods Sixty specific pathogen-free (SPF) Kunming mice (18-22 g,half male and half female) were divided into 3 groups by random number table method,20 mice in each group.One group was administered with recombinant MS-TSOL18 vaccine,one group was administered with MS as control,and one group was administered phosphate buffer saline (PBS) as control.Kunming mice were vaccinated once every two weeks for 2 times through intragastric administration.At 0,2,4,6,and 8 weeks after immunization,blood sample was collected and serum was separated.The levels of IgG and IgG2a in serum were detected using enzyme linked immunosorbent assay (ELISA).The spleen was separated to prepare spleen suspension.The proliferation of spleen lymphocyte with the stimulation of a specific antigen was detected by CCK-8.The levels of interleukin (IL)-2 and IL-4 in spleen cell culture supernatant with the stimulation of a specific antigen were detected by ELISA.Results Compared with MS control group (IgG:0.160 ± 0.019,0.187 ± 0.038,0.193 ± 0.050,0.170 ± 0.005;IgG2a:0.213 ± 0.010,0.198 ± 0.012) and PBS control group (IgG:0.159 ± 0.015,0.184 ± 0.029,0.191 ± 0.025,0.165 ± 0.018;IgG2a:0.198 ± 0.032,0.178 ± 0.025),the levels of specific IgG (0.310 ± 0.034,0.391 ± 0.029,0.443 ± 0.030,0.373 ± 0.021) in recombinant MS-TSOL18 vaccine group increased at 2,4,6,and 8 weeks after immunization (P ＜ 0.05),the levels of specific IgG2a (0.446 ± 0.056,0.339 ± 0.026) in recombinant MS-TSOL18 vaccine group increased at 6 and 8 weeks after immunization (P ＜ 0.05),and reached the highest level by the 6th week.After antigen stimulation,compared with MS and PBS control groups,the levels of spleen lymphocyte proliferation increased at 2,4,6,and 8 weeks after immunization (P ＜ 0.05),and reached the highest level by the 6th week.After antigen stimulation,compared with MS and PBS control groups,the levels of IL-2 and IL-4 in spleen lymphocyte culture supernatant increased at 2,4,6,and 8 weeks after immunization (P ＜ 0.05),and reached the highest level by the 6th and 4th weeks,respectively.Conclusion The recombinant MS-TSOL18 vaccine of Taenia solium might induce mice to produce humoral and cellular immune responses.

Objective To evaluate the efficacy of the bacterial lysate Broncho-vaxom on chronic obstructive pulmonary disease(COPD)in elderly patients.Methods A total of 150 patients with stable COPD were randomized into the Broncho-vaxom group(n=78)treated with Broncho-vaxom as add-on to conventional treatment and the control group(n=72)treated with conventional treatment.The number of acute exacerbation of COPD per patient per year,quality of life,lung function,T cell subsets were evaluated for the therapeutic effects.Results After one year of treatment,the number of acute exacerbation per patient per year was lower in Broncho-vaxom group than in control group[(1.44 ± 1.05) times/y vs.(1.82 ± 0.61) times/y,t =2.754,P =0.007].The proportions of acute exacerbationfree patients were higher in Broncho-vaxom group than in control group(20.5％ or 16/78 vs.4.2％ or 3/72,x2 =9.043,P =0.003).There were significant differences in the forced vital capacity(FVC),forced expiratory volume in one second(FEV1),and forced expiratory volume in one second/predicted value ratio(FEV1 ％)between the two groups at 5 different time points(before,at 3,6,9 and 12 months after treatment) (Broncho-vaxom group:F =90.819,50.674 and 51.233,P =0.000;control group:F =84.928,90.654 and 86.117,P =0.000).After treatment,the symptom scores were lower in Broncho-vaxom group than in control group,and the ratios of CD4/CD8 T-cell were increased and level of CD8 T-cell was decreased in Broncho-vaxom group(all P ＜ 0.05).The rate of adverse reactions had no significant difference between Broncho-vaxom group and control group(3/78 or 3.8％ vs.2/ 72 or 2.8％,x2 =0.132,P=0.717).Conclusions The oral administration of Broncho-vaxom for six months can modulate and enhance immune functions,significantly decrease acute exacerbation frequency,improve quality of life and delay the deterioration in lung function in elderly patients with stable COPD.

Objective To formulate targeted nursing measures and prevent or reduce falls of inpatients through the analysis of related risk factors of falls of spinal degenerative disease patients. Methods Review and analysis falls of inpatients happened between January 2015 and April 2016 in our department and find out that all kinds of dangerous reasons lead to falls. All inpatients risk assessments are evaluated with Morse Fall Scale on admission and during hospitalization. The grading standard of patient safety events was used in the classification of fall outcomes. Results A total of 13 inpatients were all high risk patients by Morse Fall Scale. Risk factors of falls:periods between 0:00 and 2:00, 6:00 and 8:00am, 18:00 and 20:00, in 3 days and 2 weeks after admission; combined diseases such as cardiovascular disease, diabetes, osteoarthritis of the knee; accompanied by muscle strength, muscle tension abnormalities. use related drugs;need of accompany or assisted walking;lack of safety awareness;slippery ground, compressed activity space and lack of light. Conclusions Falls of inpatients with spinal degenerative diseases are caused by the joint action of physiological difference, disease, medicine and weak safety consciousness of inpatients and caregivers, with specialty and occurrence in a certain time. Nursing measures can help to avoid and reduce the falls of inpatients.

Objective To evaluate the regulatory role of azithromycin-induced persistent Chlamydia trachomatis (Ct) infection in the apoptosis of Hela229 cells.Methods Hela229 cells were firstly co-cultured with Ct for 22 hours,and then cultured with Dulbecco's modified Eagle's medium (DMEM) containing 0.08 mg/L azithromycin for 26 hours to establish a cell model of persistent Ct infection (persistent infection group).These infected Hela229 cells cultured with azithromycin-free DMEM served as a cell model of acute Ct infection (acute infection group).After 48-hour infection with Ct,azithromycin was removed,and infected Hela229 cells in the above 2 groups were successively cultured with DMEM for the resurgence of Ct.Immunofluorescence assay and electron microscopy were performed to verify the persistent Ct infection model.The Hela229 cells in the persistent infection group and acute infection group as well as uninfected Hela229 cells (control group) were treated with staurosporine (STS) for 4 hours to induce the apoptosis,and then cell apoptosis was detected by Hoechst 33258 staining,annexin V/propidium iodide staining and flow cytometry.Results After the treatment with azithromycin,atypical inclusions with aberrant reticulate bodies appeared in the Ct-infected cells.After removing azithromycin,cells were cultured until 96 hours after infection,and infectious elementary bodies reappeared in the Ct inclusions.After the treatment with STS,Hoechst staining showed that there was loose chromatin in the persistently infected cells,while chromatin condensation was observed in the uninfected cells.After 24-hour infection with Ct and 4-hour induction with STS,the apoptosis rate was significantly higher in the persistent infection group (45.567％ ± 2.631％) than in the acute infection group (38.567％ ± 1.701％,t =2.686,P =0.028),but significantly lower in the persistent infection group than in the uninfected group (69.800％ ± 2.835％,t =8.187,P ＜ 0.001).After 48-hour infection with Ct and 4-hour induction with STS,there was a significant difference in the apoptosis rate between the persistent infection group (46.700％ ± 5.257％) and acute infection group (61.767％ ± 1.815％,t =5.781,P ＜ 0.001),as well as between the persistent infection group and the uninfected group (68.667％ ± 3.156％,t =7.421,P ＜ 0.001).Conclusion This study showed that azithromycin-induced persistent Ct infection regulated the apoptosis of host cells,and this effect lasted 48 hours.

We expressed B cell epitopes of dengue virus envelope protein and NS1 protein in prokaryotic cells,and purified and evaluated for its serological activities.A recombinant multi-epitope chimeric gene named rE including eight B cell epitopes was connected by linker peptide (EAAAK)2 and cloned into prokaryotic expression vector pET-28a(+),and transformed into E.coli BL21(DE3) cells for expression under induction of IPTG.The expressed recombinant protein was purified with 6× His purification media,and identified by SDS-PAGE and Western blot,and its antigenicity was analyzed by using an indirect ELISA assay.The recombinant expression vector pET28a-rE was constructed and expressed in BL21 (DE3) successfully,but the recombinant proteins mainly appeared as inclusion bodies.The target protein was obtained with high purity through the purification of affinity chromatography.SDS-PAGE and Western blot analysis showed that the molecular weight of fusion protein was in the expected line.The established indirect ELISA has high accuracy.This recombinant peptide antigen expressed in E.coli has good potential for serum testing.