Objective:To discuss whether safranal affects the proliferation,migration,and phenotypic transformation of the vascular smooth muscle cells(VSMCs)in a high-glucose environment and to clarify the function of safranal in the prevention and treatment of diabetic(DM)vascular complications.Methods:The SD rats were selected as experimental subjects;primary VSMCs were cultured from rat thoracic aortas and divided into control group,25 mmol·L-1 high glucose(HG)group,HG+20 μmol·L-1 safranal group,HG+40 μmol·L-1 safranal group,and HG+80 μmol·L-1 safranal group.The cells in control group received no treatment;the cells in 25 mmol·L-1 HG group were pretreated with 25 mmol·L-1 HG;the cells in HG+20,40,and 80 μmol·L-1 safranal groups were further treated with 20,40,and 80 μmol·L-1 safranal respectively for 48 h on the basis of 25 mmol·L-1 HG group.Cell counting kit-8(CCK-8)method was used to determine the appropriate concentration of safranal and detect the viabilities of the VSMCs in various groups;cell scratch healing assay was used to detect the scratch healing rates of the VSMCs in various groups;Transwell chamber assay was used to detect the numbers of the migration VSMCs in various groups;immunofluorescence method was used to detect the fluorescence intensities of alpha-smooth muscle actin(α-SMA)and rabbit anti-osteopontin(OPN)in the VSMCs in various groups;Western blotting method was used to detect the expression levels of OPN,α-SMA,and proliferating cell nuclear antigen(PCNA)in the VSMCs in various groups.Results:Under microscope,on the 4th day of in vitro culture,the spindle-shaped or triangular cells crawled out from the edge of the thoracic aorta tissue blocks,with long spindle being the most common morphology.On the 14th,the cells gradually covered the bottom of the dish;when cell density reached 80%-90%,the characteristic"hills and valleys"growth pattern appeared.Third-generation cells were taken for immunofluorescence identification;immunofluorescence staining with VSMC-specific marker α-SMA showed positive expression of α-SMA protein in the primarily cultured VSMCs.The CCK-8 assay results showed that compared with control group,the cell viability of the cells in 160 μmol·L-1 safranal group was significantly decreased(P<0.01),indicating toxic damage to the cells.Under the conditions of safranal concentrations at 20,40,and 80 μmol·L-1 respectively,after 48 h intervention on VSMCs,no significant adverse effect on cell viability was observed;considering both the effect and toxicity of safranal,these three concentrations were used in subsequent cell experiments.After 48 h intervention,compared with control group,the activity of the VSMCs in 25 mmol·L-1 HG group was increased(P<0.001);compared with 25 mmol·L-1 HG group,the activities of the VSMCs in HG+20,40,and 80 μmol·L-1 safranal groups were gradually decreased(P<0.05).The cell scratch healing assay and Transwell assay results showed that after 48 h intervention,the scratch healing rate of the VSMCs in 25 mmol·L-1 HG group was significantly higher than that in control group(P<0.01),and the number of transmembrane cells through the Transwell chamber was significantly increased(P<0.05);compared with 25 mmol·L-1 HG group,the scratch healing rates of the VSMCs in HG+20,40,and 80 μmol·L-1 safranal groups were gradually decreased(P<0.05),and the number of transmembrane cells was decreased(P<0.05).The immunofluorescence staining results showed that compared with control group,the fluorescence intensity of α-SMA protein in the VSMCs in 25 mmol·L-1 HG group was significantly weakened(P<0.001),while the fluorescence intensity of OPN protein was significantly enhanced(P<0.001);compared with 25 mmol·L-1 HG group,the fluorescence intensities of α-SMA protein in the VSMCs in HG+20,40,and 80 μmol·L-1 safranal groups were gradually increased(P<0.05),and the fluorescence intensities of OPN were gradually weakened(P<0.05).The Western blotting method results showed that compared with control group,the expression level of α-SMA protein in the VSMCs in 25 mmol·L-1 HG group was decreased(P<0.05),and the expression levels of PCNA and OPN proteins were increased(P<0.01);compared with 25 mmol·L-1 HG group,the expression level of α-SMA protein in the VSMCs in HG+20,40,and 80 μmol·L-1 safranal groups were increased(P<0.05),and the expression levels of PCNA and OPN proteins were decreased(P<0.05).Conclusion:Safranal can inhibit the proliferation,migration,and phenotypic transformation of the VSMCs induced by high glucose.

Objective: To explore the active components, key targets, and mechanism of action of turnip in alleviating pulmonary fibrosis(PF) based on network pharmacology and animal experiments. Methods: The active components and targets of Brassica rapa L. were screened using the traditional Chinese medicine systems pharmacology database and analysis platform database, and PF-related targets were obtained from disease databases such as online mendelian inheritance of man(OMIM) and DrugBank. The intersection targets were used to construct a protein-protein interaction(PPI) network to identify core targets, followed by gene oncology(GO)/Kyoto encyclopedia of genes and genomes(KEGG) pathway enrichment analysis. In the animal experiments, a bleomycin-induced PF mouse model was established. Pathological changes in lung tissue were evaluated using HE and Masson staining. qRT-PCR was used to detect the mRNA expression of tumor necrosis factor-α(TNF-α), phosphatidylinositol 3-kinase(PI3K), and akstrain transforming 1(AKT1), and immunofluorescence staining was used to measure the protein expression of TNF-α, PI3K, and AKT1. Results: The 68 active components identified in Brassica rapa L. may regulate PI3K-Akt signaling pathway by acting on 89 potential targets such as TNF-α and AKT1. The results of animal experiments showed that polysaccharide of Brassica rapa L.(BRPs) could significantly reduce the degree of bleomycin induced pulmonary fibrosis in mice; HE and Masson staining of lung tissue showed that compared with the model group, the damage of alveolar structure, the infiltration of inflammatory cells and the deposition of collagen fibers in the BRPs treatment group were significantly reduced. Further mechanism studies showed that BRPs could significantly down-regulate the mRNA and protein expression levels of TNF-α, PI3K and AKT1 in lung tissue of pulmonary fibrosis mice. Conclusion Brassica rapa L. can synergistically alleviate pulmonary fibrosis through “multi-component, multi-target and multi-channel” approach; BRPs is one of the main active components, and plays an anti-fibrosis role by inhibiting TNF-α/PI3K Akt signaling pathway.

Objective:To understand the dilemma of active management in young and middle-aged postoperative patients with lumbar disc herniation, in order to provide reference for the formulation of active intervention program of self-health management meeting the needs of patients.Methods:Using the phenomenological research method, a total of 16 middle-aged and young patients with lumbar disc herniation were selected for semi-structured in-depth interviews, and the themes were analyzed and extracted by Colaizzi 7-step analysis method.Results:Three themes and nine sub-themes were extracted namely, inadequate disease response capacity and response resources (lack of disease symptom management capacity, limited access to disease management information, difficult use of disease management information, difficulty in maintaining positive self-management for a long time (forced to stop behavior change due to stress, lack of motivation to achieve good behavior change, doubts about the effectiveness of self-management) and multi-dimensional negative emotions (contrast between maintaining independence and dependence on others, fear of being misunderstood, fear of relapse) .Conclusions:Medical staff should provide comprehensive health information services for young and middle-aged postoperative patients with lumbar disc herniation, improve the out-of-hospital follow-up mechanism with the help of information technology, further improve family and social support strategies, reduce the difficulties faced in the process of active management and achieve long-term active disease management.

DNMT3A encodes a DNA methyltransferase involved in development, cell differentiation, and gene transcription, which is mutated and aberrant-expressed in cancers. Here, we revealed that loss of DNMT3A promotes malignant phenotypes in lung cancer. Based on the epigenetic inhibitor library synthetic lethal screening, we found that small-molecule HDAC6 inhibitors selectively killed DNMT3A-defective NSCLC cells. Knockdown of HDAC6 by siRNAs reduced cell growth and induced apoptosis in DNMT3A-defective NSCLC cells. However, sensitive cells became resistant when DNMT3A was rescued. Furthermore, the selectivity to HDAC6 inhibition was recapitulated in mice, where an HDAC6 inhibitor retarded tumor growth established from DNMT3A-defective but not DNMT3A parental NSCLC cells. Mechanistically, DNMT3A loss resulted in the upregulation of HDAC6 through decreasing its promoter CpG methylation and enhancing transcription factor RUNX1 binding. Notably, our results indicated that HIF-1 pathway was activated in DNMT3A-defective cells whereas inactivated by HDAC6 inhibition. Knockout of HIF-1 contributed to the elimination of synthetic lethality between DNMT3A and HDAC6. Interestingly, HIF-1 pathway inhibitors could mimic the selective efficacy of HDAC6 inhibition in DNMT3A-defective cells. These results demonstrated HDAC6 as a HIF-1-dependent vulnerability of DNMT3A-defective cancers. Together, our findings identify HDAC6 as a potential HIF-1-dependent therapeutic target for the treatment of DNMT3A-defective cancers like NSCLC.

Objective:To explore the range of medical reference value of thyroid hormone of pregnant women in early pregnancy in Jinan after adjustment of salt iodine content, so as to provide basis for clinical diagnosis and treatment of thyroid diseases of pregnant women in early pregnancy.Methods:A prospective study was conducted in 560 pregnant women in early pregnancy (0-13 weeks) who had underwent prenatal examination and thyroid function testing in Shandong Provincial Maternal and Child Health Care Hospital from January to December 2018. At the same time, 100 healthy non-pregnant women who were examined in the Health Examination Center of Shandong Provincial Maternal and Child Health Care Hospital were selected as the control population. Urine samples of pregnant women in early pregnancy were collected, and urinary iodine content was detected by arsenic cerium catalytic spectrophotometry. Venous blood samples were collected from pregnant women in early pregnancy and control population, the serum levels of free triiodothyronine (FT 3), free thyroxine (FT 4) and thyroid stimulating hormone (TSH) were detected by electrochemiluminescence (ECL), and the medical reference values of FT 3, FT 4 and TSH were established. Results:The median urinary iodine of pregnant women in early pregnancy was 162.21 μg/L, which was in the appropriate level of iodine. After the adjustment of salt iodine content, the medical reference values of FT 3, FT 4 and TSH in pregnant women in early pregnancy were 3.86-6.15 pmol/L, 12.56-22.16 pmol/L and 0.01-3.48 mU/ L, respectively; and the medical reference values of FT 3, FT 4 and TSH in control population were 3.55-6.05 pmol/L, 9.93-20.58 pmol/L and 0.54-5.92 mU/L, respectively. Conclusions:The iodine nutrition of pregnant women in early pregnancy in Jinan is appropriate after the adjustment of salt iodine content. The medical reference values of FT 3, FT 4 and TSH in pregnant women in early pregnancy are different from those in the healthy non-pregnant women. Therefore, it is of clinical significance to establish a medical reference value range of thyroid hormone for pregnant women in early pregnancy.

Objective: To explore the effect of PDCA nursing model combined with token reward on the compliance and negative emotions of children with hypospadias. Methods: A total of 120 children with hypospadias who were admitted to Children′s hospital affiliated to Zhengzhou University from February 2017 to December 2017 were selected as study subjects. According to the time of admission, they were divided into observation group and control group, with 60 cases in each group. The control group was given routine nursing care, the observation group was given PDCA nursing mode combined with token rewards intervention on the basis of routine care. The self-made children's behavioral compliance assessment table was used to compare the compliance of the two groups of children. Achenbach Child Behavior Check List (CBCL) scores were performed before and after the intervention. The hospitalization time and the incidence of complications were calculated. Results: There was no significant difference in CBCL scores between the two groups at admission (t=0.965, P=0.102). The scores of compliance and CBCL scores of the observation group were (43.12±3.99) points and (17.62±7.30) points respectively, the scores of control group were (30.67±4.53), (24.23±6.67), the difference was statistically significant (t=15.977, 5.184, P<0.01). The hospitalization time and complication rate of the observation group were (18.73±0.84)d, 15.00% (9/60), which were significantly lower than (20.92±1.90) d, 31.67% (19/60) of the control group, and the difference was statistically significant (t=8.166, χ²=4.658, P<0.01 or 0.05). Conclusions The PDCA nursing model combined with token reward can improve the compliance of children with hypospadias and improve their negative emotions.

Objective: To explore the subunit strategy for perineal defect reconstruction and flap selection. Methods: This is a respective study of 21 patients, with perineal defect, during January 2008 to December 2018. All patients were admitted to the fifth section of Burn and Plastic Surgery in the Fourth Medical Center of the People′s Liberation Army General Hospital. There were 10 males and 11 females, aged from 4 to 68 years old, with the mean age of 26.4 years. The causes of injury included burn (n=11), trauma (n=2), Paget′s disease (n=2), Brown′s disease (n=2), perineal squamous cell carcinoma (n=3)and hemangioma (n=1). The perineum is divided into 4 subunits, according to the anatomical structure: a front area monsveneris or pubic symphysis, 2 middle areas (labia or scrotum) and a posterior area (anal). The defects ranged 23 cm×11 cm-5 cm×3 cm after perineal lesions were removed. Appropriate flaps were selected based on tissue defect. Results: Nine patients were repaired with superficial inferior epigastric artery flap, 3 patients were repaired with superficial circumflex iliac artery flap, and 2 patients were repaired with combined superficial inferior epigastric artery flap and superficial circumflex iliac artery island flap. Internal pudendal arterial perforator flap was performed in 5 patients, and anterolateral thigh perforator flap in 2 patients. The size of flap was 25 cm×12 cm-6 cm×3 cm. All flaps survived, and incisions were primary healing. Patients were followed up for 6 months to 9 years, with an average of 13 months. The patients were satisfied with the appearances and functions of the recipient and doner sites. Scars were concealed well. Conclusions Appropriate flap can be chosen to repair perineal defects, based on the subunit principle in perineum, in order to restore function and appearance, and achieve satisfactory clinical outcomes.

Objective: To explore the clinical application of ultra-pulsed CO₂ laser skin abrasion combined with micro-skin graft in the treatment of skin depigmentation. Methods: From January 2010 to June 2014, depigmented skin specimens were used for ultra-pulsed CO₂ laser skin abrasion treatment, at 0, 10, 20, 30, 40, 50, 60 mJ. HE stain was performed to observe the abrasion depth under the 100 times microscope view. The most appropriate parameter was set to completely remove the epidermis, and leave dermis undamaged. From January 2011 to December 2017, 16 patients with skin depigmentation, 12 males and 4 females, aged 12-47 years, were treated with super-pulsed carbon dioxide laser and autologous microdermis transplantation. At 1, 3, 6 and 12 months after the operation, patients′ photos were record. The color improvement was analyzed using Image-Pro Plus 6.0 software. Results: The laser energy for complete epidermis removal is different at different sites. It is 50 mJ for back skin, 40 mJ for face and hand skin and 30 mJ for neck skin. After one year of follow-up, there was no hypertrophy scar caused by over-abrasion in all 16 patients. Combined with the micro-skingrafting, 15 patients with skin depigmentation were completely resolved, with more than 90%color improvement rate. The improvement rate was 75% in a patient with un-uniform appearance, due to part failure of micro-skin graft. The neck movement and uncertain fixation were the reasons. Conclusions The appropriate energy parameters can control the abrasion depth to avoid the hypertrophic scar. Combined with micro-skin graft, the depigmentation area can be improved with uniform color and reliable effect.

Objective: To analyze the genotypes, amino acid vatiations and molecular epidemiological characteristics of respiratory syncytial virus (RSV) infection in pediatric patients in Gansu province for the future research. Methods: A total of 4 556 respiratory tract specimens were colleted from pediatric patients under 10 years of age in five cities in Gansu from 2012 to 2017. These specimens were tested for RSV and its subtypes.The coding region of the RSV G gene was amplified using reverse transcription-polymerase chain reaction (RT-PCR) and sequenced for RSV positive specimens. Sequences were edited using DNA Star software. Sequence alignment and phylogenetic trees were built by MEGA 6.0 software. Results: Out of 4 556 specimens, 1 135 (24.91%) were positive for RSV, totally 216 G protein sequences were obtained. RSV A isolates were clustered into three genotypes: NA1、NA3 and ON1. The nucleotides and amino-acid homology was 84.9%-100% and 77.3%-100%, respectively. The nucleotides and amino-acid homology between this study and prototype long strain was 81.2%-83.3% and 74.1%-88.0%. RSV B isolates were clustered into only BA9 one genotypes. The nucleotides and amino-acid homology was 97.7%-100% and 95.8%-100%, respectively. The nucleotides and amino-acid homology between this study and prototype CH18537 strain was 84.9%-85.7% and 77.9%-80.1%. Conclusions The genetic characteristics and the amino-acid changes were analyzed systematically using data of RSV G gene collected from 2012 to 2017 in Gansu province in this study. These data were used for analyses of the etiology, control and prevention of RSV infection.

Non-small-cell lung cancer (NSCLC), the most common type of lung cancer accounting for 85% of the cases, is often diagnosed at advanced stages owing to the lack of efficient early diagnostic tools. 5-Hydroxymethylcytosine (5hmC) signatures in circulating cell-free DNA (cfDNA) that carries the cancer-specific epigenetic patterns may represent the valuable biomarkers for discriminating tumor and healthy individuals, and thus could be potentially useful for NSCLC diagnosis. Here, we employed a sensitive and reliable method to map genome-wide 5hmC in the cfDNA of Chinese NSCLC patients and detected a significant 5hmC gain in both the gene bodies and promoter regions in the blood samples from tumor patients compared with healthy controls. Specifically, we identified six potential biomarkers from 66 patients and 67 healthy controls (mean decrease accuracy >3.2, P < 3.68E-19) using machine-learning-based tumor classifiers with high accuracy. Thus, the unique signature of 5hmC in tumor patient's cfDNA identified in our study may provide valuable information in facilitating the development of new diagnostic and therapeutic modalities for NSCLC.
5-Methylcytosine ; analogs & derivatives ; blood ; Biomarkers, Tumor ; blood ; genetics ; Carcinoma, Non-Small-Cell Lung ; blood ; diagnosis ; genetics ; Case-Control Studies ; Circulating Tumor DNA ; blood ; DNA Methylation ; Epigenomics ; Female ; Humans ; Lung Neoplasms ; blood ; diagnosis ; genetics ; Male ; Middle Aged