Objective:To analyze the effect of long noncoding RNA (lncRNA) small nucleolar RNA host gene 16 (SNHG16) on the radiosensitivity of cervical cancer cells and explore its regulatory role in the miR-455-3p/ nuclear factor-κB (NF-κB) signaling pathway.Methods:The expression levels of lncRNA SNHG16 and miR-455-3p in human normal cervical epithelial cells H8, human cervical cancer cells SiHa, and radioresistant cervical cancer cells SiHa-R were detected by real-time reverse transcription polymerase chain reaction. SiHa-R cells were transfected separately, and then given a single dose of 4 Gy X-ray irradiation and continued to be cultured for subsequent experiments. The cells in each group were named siRNA-NC, siRNA-SNHG16 (interfering lncRNA SNHG16), NC mimic, miR-455-3p mimic (overexpressing miR-455-3p), siRNA-SNHG16+inhibitor NC, and siRNA-SNHG16+miR-455-3p inhibitor groups, respectively. The survival fraction of SiHa-R cells was detected by clone formation assay. The apoptosis rate of SiHa-R cells was analyzed by flow cytometry. The expression levels of apoptotic proteins [cysteine-containing aspartate-specific protease (Caspase)-3, Caspase-9, Bax] and NF-κB signaling pathway related proteins [NF-κB p65, phosphorylated (p)-NF-κB p65, p-IκB (inhibitory protein of NF-κB)] were measured by Western blot. The targeting relationship between lncRNA SNHG16 and miR-455-3p was determined by dual luciferase reporter gene assay. Comparison among different groups was conducted by one-way ANOVA, and paired comparison was carried out by LSD t-test. Comparison between two groups was performed by t-test. Results:Compared with H8 cells, the expression levels of lncRNA SNHG16 were increased in SiHa and SiHa-R cells, and SiHa-R cells had a higher level than SiHa cells. The expression levels of miR-455-3p were decreased in SiHa and SiHa-R cells, and SiHa-R cells had a lower level than SiHa cells (all P<0.001). Compared with the siRNA-NC group, the survival fraction of SiHa-R cells in the siRNA-SNHG16 group was decreased, the radiosensitization ratio (SER) was 1.571 (>1), the apoptosis rate and levels of Caspase-3, Caspase-9, and Bax proteins were increased, while the levels of p-NF-κB p65 and p-IκB proteins were decreased (all P<0.001). lncRNA SNHG16 could target miR-455-3p. Compared with the NC mimic group, miR-455-3p level in the miR-455-3p mimic group was increased, cell survival fraction was decreased, the SER was 1.826 (>1), the apoptosis rate and the levels of Caspase-3, Caspase-9, Bax proteins were increased, and the levels of p-NF-κB p65 and p-IκB proteins were decreased (all P<0.001). Inhibition of miR-455-3p expression could weaken the effect of interfering with lncRNA SNHG16 expression on SiHa-R cell apoptosis, radiotherapy sensitivity, and NF-κB signaling pathway (all P<0.001). Conclusions:Interference with lncRNA SNHG16 expression could induce the apoptosis of cervical cancer cells and enhance their radiation sensitivity by regulating the miR-455-3p/NF-κB signaling pathway.

Objective:To analyze the effect of long noncoding RNA (lncRNA) small nucleolar RNA host gene 16 (SNHG16) on the radiosensitivity of cervical cancer cells and explore its regulatory role in the miR-455-3p/ nuclear factor-κB (NF-κB) signaling pathway.Methods:The expression levels of lncRNA SNHG16 and miR-455-3p in human normal cervical epithelial cells H8, human cervical cancer cells SiHa, and radioresistant cervical cancer cells SiHa-R were detected by real-time reverse transcription polymerase chain reaction. SiHa-R cells were transfected separately, and then given a single dose of 4 Gy X-ray irradiation and continued to be cultured for subsequent experiments. The cells in each group were named siRNA-NC, siRNA-SNHG16 (interfering lncRNA SNHG16), NC mimic, miR-455-3p mimic (overexpressing miR-455-3p), siRNA-SNHG16+inhibitor NC, and siRNA-SNHG16+miR-455-3p inhibitor groups, respectively. The survival fraction of SiHa-R cells was detected by clone formation assay. The apoptosis rate of SiHa-R cells was analyzed by flow cytometry. The expression levels of apoptotic proteins [cysteine-containing aspartate-specific protease (Caspase)-3, Caspase-9, Bax] and NF-κB signaling pathway related proteins [NF-κB p65, phosphorylated (p)-NF-κB p65, p-IκB (inhibitory protein of NF-κB)] were measured by Western blot. The targeting relationship between lncRNA SNHG16 and miR-455-3p was determined by dual luciferase reporter gene assay. Comparison among different groups was conducted by one-way ANOVA, and paired comparison was carried out by LSD t-test. Comparison between two groups was performed by t-test. Results:Compared with H8 cells, the expression levels of lncRNA SNHG16 were increased in SiHa and SiHa-R cells, and SiHa-R cells had a higher level than SiHa cells. The expression levels of miR-455-3p were decreased in SiHa and SiHa-R cells, and SiHa-R cells had a lower level than SiHa cells (all P<0.001). Compared with the siRNA-NC group, the survival fraction of SiHa-R cells in the siRNA-SNHG16 group was decreased, the radiosensitization ratio (SER) was 1.571 (>1), the apoptosis rate and levels of Caspase-3, Caspase-9, and Bax proteins were increased, while the levels of p-NF-κB p65 and p-IκB proteins were decreased (all P<0.001). lncRNA SNHG16 could target miR-455-3p. Compared with the NC mimic group, miR-455-3p level in the miR-455-3p mimic group was increased, cell survival fraction was decreased, the SER was 1.826 (>1), the apoptosis rate and the levels of Caspase-3, Caspase-9, Bax proteins were increased, and the levels of p-NF-κB p65 and p-IκB proteins were decreased (all P<0.001). Inhibition of miR-455-3p expression could weaken the effect of interfering with lncRNA SNHG16 expression on SiHa-R cell apoptosis, radiotherapy sensitivity, and NF-κB signaling pathway (all P<0.001). Conclusions:Interference with lncRNA SNHG16 expression could induce the apoptosis of cervical cancer cells and enhance their radiation sensitivity by regulating the miR-455-3p/NF-κB signaling pathway.

Objective To investigate the long non-coding RNA(lncRNA) MRAK08838 regulates macrophage function to influence the development of asthmatic airway inflammation. Methods MRAK088388 gene knockout (MRAK088388^-/-) mouse model was prepared and allergic asthma was induced by dust mite protein Dermatophagoides farinae 1 (Der f1). The mice were sacrificed after 28 days of modeling, and serum was collected to measure IgE and IgG. The FinePointe RC system was used to measure airway hyperresponsiveness and evaluate lung function in mice. Lung tissue was taken for HE staining, and periodic acid-Schiff (PAS) staining was used to evaluate inflammatory infiltration and mucus secretion in mouse lungs. Fluorescence quantitative PCR was used to detect the expression level of lncRNA MRAK08838 in bronchoalveolar lavage fluid (BALF) cells and lung tissue of asthmatic mice. ELISA was used to detect the levels of inflammatory cytokines IFN-γ, IL-4, IL-5, IL-13, IL-10 and IL-17A. Flow cytometry was used to evaluate the phenotype of macrophages in BALF and lung tissue, as well as the proportion of neutrophils, eosinophils, and alveolar macrophages. The changes of the above indicators were detected in mice by adoptive transfer of bone marrow-derived macrophages (BMDM). Results Under the challengle of Der f1, MRAK088388^-/- mice showed reduced allergic airway inflammation, including reduced eosinophils in BALF and reduced production of IgE and IgG1. In addition, Der f1-treated MRAK088388^-/- mice had fewer M2 macrophages than wild-type asthmatic mice. Wild-type mouse BMDM (M0) and Der f1-treated MRAK088388^-/- mice also showed mild inflammatory response. Conclusion Knockout of MRAK088388 alleviates airway inflammation in asthmatic mice by inhibiting M2 polarization of airway macrophages.
Animals ; Mice ; Mice, Knockout ; RNA, Long Noncoding/genetics* ; Asthma/genetics* ; Macrophages ; Immunoglobulin E

【Objective】 To investigate the association between the long-stranded non-coding ribonucleic acid (lncRNA) MRAK088388 and allergic asthma in children. 【Methods】 A total of 15 healthy children and 15 children with asthma were monitored for disease progression over a 2-year period. Blood samples were collected from patients during the chronic phase of the disease for lncRNA/mRNA expression microarray analysis. Competing endogenous RNA networks (MRAK088388/miR-30a/ATG5) were identified by bioinformatics analysis. In vitro cultured bronchial epithelial (16HBE) cells were used to quantify gene and associated protein expression levels by real-time fluorescent quantitative polymerase chain reaction (qPCR) and protein blotting, respectively. Cell Counting Kit-8 and transwell assays were used to assess the proliferation and migration of 16HBE cells and verify the effects of MRAK088388, miR-30a and ATG5 on asthma. 【Results】 Six lncRNA-miRNA-mRNAs were identified by correlation analysis. By qRT-PCR analysis, MRAK088388/miR-30a/ATG5 was selected to construct the ceRNA network in this study. mRAK088388 and ATG5 expressions were high in the peripheral blood of children with asthma, while the expression of miR-30a was low (P<0.05). The expression level of E-cadherin was significantly higher in 16HBE cells after si-MRAK088388+TGF-β1 group, while the expression levels of Vimentin and α-SMA were significantly lower (P<0.05), indicating that knockdown of MRAK088388 inhibited the epithelial mesenchymal transition in 16HBE cells. Compared with si-NC+ TGF-β1 group, the cell morphology of si-MRAK088388+TGF-β1 group was similar to that of the control group, indicating that MRAK088388 knockdown attenuated TGF-β1-induced cell morphological changes; in addition, MRAK088388 knockdown inhibited TGF-β1-induced proliferation and migration of 16HBE cells. MRAK088388 was confirmed by qPCR and protein blotting to promote the progression of childhood asthma by targeting the miR-30a/ATG5 axis. 【Conclusion】 Childhood asthma is associated with the MRAK088388/miR-30a/ATG5 axis, and MRAK088388 is involved in the process of childhood allergic asthma by negatively regulating miR-30a expression and regulating elevated ATG5 expression levels to affect bronchial epithelial cell mesenchymal transition, proliferation, and migration.

Objective : To study the effect of ubiquinol⁃cytochrome C reductase subunit 2 (QCR2) on cell cycle arrest of cervical cancer SiHa cell line , and to explore related mechanisms. Methods : The log⁃phase SiHa cells were taken , and QCR2 siRNA and control siRNA were transfected into SiHa cells by liposome transfection , which were set as QCR2 siRNA group and control siRNA group , and untreated cells were set as a blank group. qRT⁃PCR and Western blot were used to determine the relative expression of QCR2 , p53 mRNA and protein. Propidium iodide(PI) staining was used to determine cell cycle. The cells in the QCR2 siRNA group were taken , and were intervened with a final concentration of 50 nmol/L ubiquitin⁃proteasome inhibitor PS341 as the QCR2 siRNA + PS341 group. In addition , the cells in the QCR2 siRNA group were intervened with the same amount of normal saline (NS) and set as the QCR2 siRNA + NS group. Western blot was used to determine the relative expression of p53 protein. The immunoprecipitation test was used to determine the level of p53 ubiquitination. Results : Observed under a fluorescence microscope , the transfection efficiency of QCR2 siRNA group and control siRNA group were both > 80% . Compared with the blank group and control siRNA group , the relative expression of QCR2 mRNA and protein in the QCR2 siRNA group decreased (P < 0. 05) , the proportion of G0/G1 increased (P < 0. 05) , and the proportion of S , G2/M decreased (P < 0. 05) . There was no significant difference in the relative expression of p53 mRNA between the groups. Compared with the blank group and control siRNA group , the relative expression of p53 protein in the QCR2 siRNA group increased (P < 0. 05) . Compared with the blank group and QCR2 siRNA + PS341 group , the relative expression of p53 protein in the QCR2 siRNA group and QCR2 siRNA + NS group increased (P < 0. 05) . Compared with the blank group and control siRNA group , the degree of p53 ubiquitination in the QCR2 siRNA group was reduced ( P < 0. 05 ) . Conclusion Silencing QCR2 can block SiHa cells in G0/G1 phase. Its mechanism may be related to the inhibition of p53 ubiquitination and the increase of its protein expression.

Objective: To observe the effects of minimally invasive tangential excision in treating deep partial-thickness burn wounds on trunk and limbs in pediatric patients in the early stage post burn. Methods: Clinical data of 40 children with deep partial-thickness burn wounds on trunk and limbs, admitted to our burn ward from January 2016 to June 2017, conforming to the study criteria, were retrospectively analyzed. They were divided into conventional treatment group (CT, n=19) and minimally invasive tangential excision group (MITE, n=21) according to the different treatments. The patients in group CT were treated with eschar-reserving therapy firstly. When tangential excision was performed, the roller knife was used, and no necrotic tissue left on the wound bed was considered the proper depth of excision. Razor-thickness skin grafting was performed to cover the wound when adipose tissue exposed markedly after tangential excision. Dressing change was performed within 48 h after the operation and repeated every 2 days. Unhealed wounds were covered by razor-thickness skin grafting. The patients in group MITE were treated with tangential excision in the early stage post burn. The tangential excision was operated with electric dermatome, and the thickness was set at 0.1 mm to excise the surface of eschar until the sporadic punctate hemorrhage on wound surface was observed and some necrotic tissue was left on the wound bed. Porcine acellular dermal matrix was applied after tangential excision. The first dressing change was often performed about 1 week after the operation. Razor-thickness skin grafting was performed to cover the unhealed wounds. The length of wound healing, high fever, antibiotic usage, and hospital stay, times of later operation, and hospitalization expenses of patients in the 2 groups were recorded. The excisional eschar and wound bed tissue of patients in group MITE were harvested for pathological observation. Data were processed with t test and Fisher′s exact probability test. Results: (1) There were no statistically significant differences in length of high fever and length of hospital stay and hospitalization expenses between patients in the 2 groups (t=-1.67, -1.93, 0.31, P>0.05). The lengths of wound healing [(24.8±2.5) d] and antibiotic usage [(4.4±0.7) d] of patients in group MITE were significantly shorter than those in group CT [(33.3±2.5) and (7.0±0.7) d, t=-2.44, -2.44, P<0.05], and times of later operation of patients in group MITE [(0.29±0.14) times] were significantly less than those in group CT [(0.79±0.21) times, t=-2.03, P<0.05]. (2) The thickness of the excisional eschar of patients in group MITE was about 150 μm. The eschar has epidermis and upper dermis. Some necrotic tissue was left on the wound bed. Conclusions The treatment for pediatric deep partial-thickness burn wounds on trunk and limbs with minimally invasive tangential excision using electric dermatome in the early stage post burn can accelerate wound healing, shorten length of antibiotic usage, and reduce times of later operations.

We report the results of detecting M. lepraespecific antigen--phenolic glycolipid I (PGL--I) with modified M-Dot-ELISA in sera from 75 cases of household contacts (HC)of leprosy patients, which were all seropositive by Gelatin Particle Agglutination Test (MLPA)and ELISA. The results indicate that: (1 ) 16/75 (21. 3 % ) are antigen positive. The rates of positivity in HC of MB patients are much higher than those in HC of PB patients,the difference (P