Objective: To investigate the expression，prognosis and biological role of hepatocyte nuclear factor 4A (HNF4A) in gastric cancer，and to study its effect on the proliferation of gastric cancer cells． Methods: Tumor Immune Estimation Resource 2. 0 (TIMER2. 0) and Gene Expression Profiling Interactive Analysis ( GEPIA2) databases were used to analyze the relative expression levels of HNF4A in gastric cancer and normal tissue，KM Plotter was used to analyze the correlation between the expression level of HNF4A and the survival rate of gastric cancer patients，TISIDB database and R language (4. 1. 2) were used to analyze whether HNF4A was involved in the immune regulation process of gastric cancer．cBioPortal database was used to analyze the mutations of HNF4A in gastric cancer，GSEA 4. 2 was used to analyze the functional enrichment of HNF4A，and LinkedOmics database was used to predict the genes that might be regulated by HNF4A．The relative expression of HNF4A in gastric cancer and adjacent tissues was detected by qRT-PCR , Western blot and immunohistochemistry (IHC) ．The proliferation and cell cycle of gastric cancer cells were analyzed by CCK-8，EdU，colony forming assay and flow cytometry. Results : The expression of HNF4A increased in gastric cancer tissues (P ＜0. 05) ，and the overall survival rate of gastric cancer patients with high HNF4A expression was worse (P＜0. 001) ．HNF4A was mainly missense mutated in gastric cancer．Immune cell infiltration showed that HNF4A was associated with B lymphocytes，CD8 + T cells， neutrophils，macrophages and dendritic cells ( all P ＜0. 001) ． HNF4A was also associated with tumor mutation burden (r = 0. 28，P＜0. 0001) and microsatellite instability (r = 0. 13，P＜0. 01) ．After knockdown of HNF4A， cell proliferation ability was significantly inhibited ，and cell cycle was arrested at G0 / G1 phase． Conclusion HNF4A expression significantly increased in gastric cancer tissues，which is associated with poor prognosis ，and may also be involved in immune regulation． Knockdown of HNF4A can inhibit the proliferation of gastric cancer cells.

In recent years, with the advancement of prevention and treatment concepts and diagnostic technology, the diagnosis rate of deep vein thrombosis (DVT) has increased year by year. In order to prevent the occurrence of DVT, a simple, economical, effective and practical exercise method, namely ankle pump exercise, is widely used. However, in clinical practice, there is no uniform standard for the method and duration of ankle pump exercise. This article elaborates on the overview, exercise methods, applicable population, clinical effectiveness and health education of ankle pump exercise, and summarizes the related research of ankle pump exercise, so as to provide the nursing staff with effective health guidance and provide evidence for DVT prevention.

OBJECTIVE: To explore the effects of nano-silicon dioxide( SiO_2) on the survival and poly( ADP-ribose)polymerase-1( PARP-1) expression in human bronchial epithelial cells( 16 HBE cells). METHODS: i) The 16 HBE cells were treated with nano-SiO_2 at concentrations ranging from 0 to 100 mg/L for 24. 0 hours,and CCK-8 assay was used to examine cell viability. ii) The 16 HBE cells were divided into 6 groups: solvent control group( equal volume solvent treatment),micro-SiO_2 control group( treated with 20 mg/L micro-SiO_2),5,10,and 20 mg/L nano-SiO_2 groups( treated with the corresponding final dose of nano-SiO_2),and curcumin group. The curcumin group was given pretreatment with curcumin at a final concentration of 10 μmol/L for 2. 0 hours followed by treatment with a final concentration of 20 mg/L of nano-SiO_2. Cells in each group were harvested at time points of 4. 0,12. 0 and 24. 0 hours after treatment. The relative expression of PARP-1 mRNA and protein in 16 HBE cells was detected by quantitative real-time polymerase chain reaction and Western blotting respectively. RESULTS: i) The survival of 16 HBE cells decreased with increasing nano-SiO_2 treatment dose,showing a dose-effect relationship( P < 0. 01). ii) The expression of PARP-1 mRNA and protein in 16 HBE cells were dose-dependently decreased after nano-SiO_2 stimulation at the 12. 0 and 24. 0 hours time points( P < 0. 01). The expression of PARP-1 mRNA and protein in 5,10,and 20 mg/L nano-SiO_2 groups decreased at the above mentioned time points( P < 0. 05),compared with the solvent control group at the same time points. The expression of PARP-1 mRNA and protein in 20 mg/L nano-SiO_2 group was lower than that in the micro-SiO_2 control group at the same 12. 0 and 24. 0 hours time point( P < 0. 05). The above two indexes of cells were higher in curcumin group than that of 20 mg/L nano-SiO_2 group at the 12. 0 hours time point( P < 0. 05). CONCLUSION: Nano-SiO_2 stimulation can lead to decrease survival of 16 HBE cells in a dose-dependent manner and down-regulation of PARP-1 expression may be one of the mechanisms of proliferation and inhibition of 16 HBE cells induced by nano-SiO_2. Curcumin has certain protective effect on nano-SiO_2-induced 16 HBE cell injury.

Objective: To determine the levels of vitamin D in patients with pulmonary tuberculosis in Shenzhen and identify the influencing factors of vitamin D levels and key groups of vitamin D deficiency, so as to provide a scientific basis for tuberculosis- and nutrition-related health education and promotion in Shenzhen. Methods: Patients with smear-positive pulmonary tuberculosis who were diagnosed in 2016 were selected as the research subjects. Their relevant information and blood samples were collected, and the sample pool was established according to the inclusion criteria. One hundred and twenty patients were selected based on simple random sampling, including 84 men (70.0%) and 36 women (30.0%). Blood 25-hydroxyvitamin D [25(OH)D] concentrations were measured using chemiluminescence technology. Vitamin D statuses in patients were statistically described, and vitamin D levels in patients with different characteristics were compared. Multivariate linear regression analysis was performed to identify important factors influencing vitamin D levels in patients. Results: Mean serum concentration of 25(OH)D in 120 patients was (40.2±16.0) nmol/L. There were 2 cases of vitamin D sufficiency (1.7%), 28 cases of vitamin D insufficiency (23.3%), and 90 cases of vitamin D deficiency (75.0%), of which 23 cases (19.2%) were of severe deficiency. 25(OH)D concentrations in patients with different lifestyles (indoors; indistinguishable indoors or outdoors; outdoors) were significantly different (35.3 nmol/L vs. 40.6 nmol/L vs. 49.5 nmol/L, F=8.274, P<0.001). Mean concentration of 25(OH)D in patients with sun exposure time of >30 min/d was higher compared to that in those with sun exposure time <30 min/d in the last month (46.4 nmol/L vs. 36.7 nmol/L, t=3.342, P=0.001). Multivariate linear regression analysis showed that the risk factors for 25(OH)D with statistical significance were Han ethnicity or not (β=-11.576, t=-1.991, P=0.049), housekeeping (including unemployment and retirement) or not (β=-6.136, t=-1.998, P=0.048), sun exposure time<30 min/d or none (β=-9.644, t=-2.829, P=0.006), body mass index (β=-2.056, t=-3.439, P=0.001) , and indoor degree of lifestyles (β=-4.419, t=-2.155, P=0.033). Conclusion Level of vitamin D is generally insufficient in patients with pulmonary tuberculosis in Shenzhen. It is necessary to strengthen health education related to vitamin D in patients with tuberculosis, especially in the high-risk population of vitamin D deficiency, such as in those with a lack of exposure to sunlight or high BMI.

Objective To evaluate the effect of micronutrient selenium on atopic dermatitis-like skin lesions in mice.Methods After 4-week feed with forages lacking selenium,40 BALB/c mice were randomly and equally divided into 4 groups:selenium deficiency group fed with forages containing 0.01 mg/kg selenium for 4 weeks,normal selenium supplementation group fed with forages containing 0.25 mg/kg selenium for 4 weeks,excessive selenium supplementation group fed with forages containing 3.00 mg/kg selenium for 4 weeks,and control group fed with forages containing 0.25 mg/kg selenium for 4 weeks.Then,atopic dermatitis-like skin lesions were induced by dinitrochlorobenzene (DNCB) in the mice in sensitized groups,including the selenium deficiency group,normal selenium supplementation group and excessive selenium supplementation group.During sensitization,the severity of dermatitis in mice was monitored.Three weeks after the sensitization,the total plasma IgE level and inflammatory cell count in whole blood were measured.Skin tissues from the back of mice were subjected to histopathological examination and the selenium level was detected.Statistical analysis was carried out by one-way analysis of variance for comparisons of IgE levels and cell counts among different groups,as well as by Pearson correlation analysis and linear regression analysis for analyzing the correlation of various indices with the selenium level.Results Six days after the sensitization,the dermatitis severity scores were significantly higher in the sensitized groups than in the control group (On day 6,8,11,13,15 and 18 after the sensitization,F =44.897,76.622,114.866,33.352,28.605 and 11.271 respectively,all P ＜ 0.01).On day 11 after the sensitization,the dermatitis severity score was significantly higher in the excessive selenium supplementation group than in the selenium deficiency group and normal selenium supplementation group (both P ＜ 0.05).Compared with the control group,obvious pathological changes of dermatitis were observed in the sensitized mice,but there was no significant difference in the number of inflammatory cells in skin lesions among the 3 sensitized groups (all P ＞ 0.05).The inflammatory cell count in whole blood and total plasma IgE level in the sensitized mice increased along with the increase of dietary selenium levels,and the excessive selenium supplementation group showed higher total plasma IgE level ([167.17 ±8.49] μg/L) compared with the selenium deficiency group ([124.78 ± 5.32] μg/L,t =3.919,P ＜ 0.05),normal selenium supplementation group ([132.61 ± 4.71] μg/L,t =3.222,P ＜ 0.05) and control group ([109.13 ± 0.79] μg/L,t =6.485,P ＜ 0.05).The selenium level in the skin tissues of sensitized mice was positively linearly correlated with the total plasma IgE level (r =0.579,P ＜ 0.001),whole-blood white blood cell count (r =0.414,P ＜ 0.05),neutrophil count (r =0.439,P ＜ 0.05),lymphocyte count (r =0.417,P ＜ 0.05)and eosinophil count (r =0.505,P ＜ 0.01).Conclusion Different dietary selenium levels showed different effects on the severity of dermatitis in mice,and more severe dermatitis occurred in the excessive selenium supplementation group.

Objective To evaluate inhibitory effect of Chlamydia trachomatis plasmid-encoded protein pORF5 on HeLa cell apoptosis induced by tumor necrosis factor-alpha(TNF-α). Methods The recombinant lentiviral expression vector containing pORF5 gene and helper plasmids were co-transfected into 293T cells to prepare the recombinant lentivirus. Then, the lentivirus particles were collected and concentrated, and used to infect HeLa cells. Flow cytometric screening identified stable pORF5-expressing HeLa (pORF5-HeLa) cells. Meanwhile, the empty plasmid was transfected into HeLa cells to prepare control HeLa cells. The two cell lines were both divided into two subgroups to be treated with 20μg/L TNF-αand fresh culture medium respectively for 6 hours. Then, Hoechst 33258 staining was performed to observe morphological changes of apoptotic cells, flow cytometry to detect cell apoptosis, real-time PCR to measure the mRNA expression of Caspase3, Bax and Bcl-2, and Western blot analysis to determine the protein expression of Bax and Bcl-2. Results After 6-hour treatment with TNF-α, Hoechst 33258 staining showed variable degrees of karyopyknosis and karyorrhexis, and highly-refractive blue apoptotic bodies in the pORF5-HeLa cells and control HeLa cells. The pORF5-HeLa cells and control HeLa cells both showed significantly higher apoptosis rate in the treated subgroup than in the untreated subgroup (pORF5-HeLa cells:35.5%± 4.5%vs. 9.5%± 1.5%, t=13.53, P<0.01;control HeLa cells:63.6%± 5.8%vs. 7.9%± 0.9%, t=32.36, P<0.01). Compared with treated control HeLa cells, treated pORF5-HeLa cells showed significant decreases in mRNA expression of Bax(72.8%)and Caspase 3(84.5%)(t = 35.29, 42.25, respectively, both P < 0.01), as well as in Bax protein expression(t = 17.58,P < 0.01), but significant increases in Bcl-2 mRNA and protein(6.8 times)expression(t = 87.12, 18.93, respectively, both P <0.01). Conclusion pORF5 plasmid protein can inhibit TNF-α-induced HeLa cell apoptosis likely by increasing the expression of anti-apoptotic protein bcl-2 and decreasing the expression of pro-apoptotic proteins Caspase-3 and Bax.

OBJECTIVETo investigate the effect of short and long term exposure to SiO2 nanoparticles on microRNA expression level in human bronchial epithelial cells(16HBE cells).

METHODSThe 16HBE cells were exposed to 5, 10, 15, 20, 25, 30 and 40 μg/ml SiO2 nanoparticles for 24 h to detect the cell viability by using CCK-8 assay. The inhibition rate of proliferation activity and half inhibitory concentration (IC50) were calculated. The 16HBE cells were exposed to 10 μg/ml SiO2 nanoparticles for 10 and 30 generations, named P10 and P30, and the control P0 was set. The cells were treated with SiO2 nanoparticles at 0, 1/4 IC50, 1/2 IC50 and IC50 concentration and μm-SiO2 at IC50 concentration for 24 h, and the control serum-free culture medium was set. Agilent miRNAs microarray chip was used to screen differentially expressed miRNAs in P10, P30 and P0 groups. The expression level of miRNA was detected by reverse transcription fluorescence quantitative polymerase chain reaction (qRT-PCR).

RESULTSThe inhibition rate of proliferation activity of 5, 10, 15, 20, 25,30,40 μg/ml group were (-3.33 ± 3.80)%, (20.40 ± 11.73)%, (39.08 ± 5.53)%, (55.10 ± 5.78)%, (66.42 ± 9.60)%, (71.67 ± 7.34)%, (81.43 ± 5.37)%, respectively; F=129.11, P<0.001. The IC50 (95%CI) was 18.35 (15.82-20.72) μg/ml. The expression level of miRNA-494-3p in P0, P10 and P30 were 1.00, 0.45 ± 0.08, 0.28 ± 0.07, respectively; F=60.77, P<0.001. miRNA-19a-3p were 1.00, 2.27 ± 0.45, 1.06 ± 0.19, respectively; F=30.05, P<0.001. miRNA-148b-3p were 1.00, 1.78 ± 0.29, 0.88 ± 0.19, respectively; F=30.23, P<0.001. Compared to control group, the expression level of miRNA-494-3p in 5, 10, 20 μg/ml SiO2 nanoparticles groups and 20 μg/ml μm-SiO2 group were 0.99 ± 0.04, 1.38 ± 0.19, 2.13 ± 0.14, 0.81 ± 0.25, respectively; F=57.03, P<0.001. miRNA-19a-3p were 0.91 ± 0.03, 1.12 ± 0.03, 0.53 ± 0.01, 0.86 ± 0.01, respectively; F=408.78, P<0.001. miRNA-148b-3p were 0.95 ± 0.02, 1.22 ± 0.00, 0.54 ± 0.02, 1.15 ± 0.04 respectively; F=264.14, P<0.001.

CONCLUSIONShort and long term exposure to SiO2 nanoparticles can affect the expression level of miRNAs in 16HBE cells. The expressions of miRNA-494-3p after long and short period exposure are different.

Cells, Cultured ; Epithelial Cells ; drug effects ; metabolism ; Humans ; MicroRNAs ; metabolism ; Nanoparticles ; chemistry ; Oligonucleotide Array Sequence Analysis ; Silicon Dioxide ; chemistry

OBJECTIVETo study the effect of silicon dioxide nanoparticles on the expression and promoter region CpG islands methylation of (Poly [ADP-ribose] polymerase 1, PARP-1) gene in human HaCaT Cell.

METHODSHaCaT Cells were treated with nm-SiO₂at 0, 2.5, 5 and 10 µg/mL and micro-SiO₂at 10 µg/ml for 24 h and DAC treatment was given at 10 µg/ml group for 48 h. Real-time PCR and western blot assay was used to detect the expression of PARP-1 mRNA and protein. BSP (Bisulfite Pyrosequence, BSP) assay was used to detect the promoter region CpG islands methylation status of PARP-1 gene.

RESULTSAfter exposure to nano-SiO₂particles, compared to CTRL group, the mRNA and protein expression of PARP-1 in micro-SiO₂and 2.5 µg/ml group unchanged, but he mRNA and protein expression of PARP-1 in 5, 10 µg/ml as well as DAC group was down-regulated and there are statistical significance between CTRL group and 5, 10 µg/ml as well as DAC group and the PARP-1 promoter region CpG islands showed methylation.

CONCLUSIONnano-SiO₂can down-regulate PARP-1 expression in HaCaT Cell and this is associated with the change in the methylation of PARP-1 gene promoter region CpG islands induced by nano-SiO₂particles.

Cell Line, Tumor ; CpG Islands ; DNA Methylation ; Humans ; Nanoparticles ; adverse effects ; Poly (ADP-Ribose) Polymerase-1 ; Poly(ADP-ribose) Polymerases ; metabolism ; Promoter Regions, Genetic ; RNA, Messenger ; metabolism ; Silicon Dioxide ; adverse effects

OBJECTIVETo establish a mechanical simulation model for studying the relationship between the characteristic frequency and feature location of the basilar membrane of the cochlea.

METHODSMacro-mechanical methods were used to simplify the details of the model. With simulation tools, the basilar membrane vibration frequency characteristics were analyzed based on the box model.

RESULTSThe basilar membrane had obvious frequency-selective properties, and the basilar membrane from the stapes was sensitive to high frequencies while the farther membrane was sensitive to low frequencies.

CONCLUSIONThe frequency characteristics of the basilar membrane of the cochlea is mainly a result of the longitudinal variations of the geometric dimensions and material properties and is not related with other structures within the cochlea corti.

Basilar Membrane ; physiology ; Cochlea ; physiology ; Computer Simulation ; Mechanics ; Models, Biological ; Vibration

Objective To establish a mechanical simulation model for studying the relationship between the characteristic frequency and feature location of the basilar membrane of the cochlea. Methods Macro-mechanical methods were used to simplify the details of the model. With simulation tools, the basilar membrane vibration frequency characteristics were analyzed based on the box model. Results The basilar membrane had obvious frequency-selective properties, and the basilar membrane from the stapes was sensitive to high frequencies while the farther membrane was sensitive to low frequencies. Conclusion The frequency characteristics of the basilar membrane of the cochlea is mainly a result of the longitudinal variations of the geometric dimensions and material properties and is not related with other structures within the cochlea corti.