Objective To investigate the relationship between circular RNA homeodomain interacting protein ki-nase 3 (circHIPK3) and the activation of rat microglia (RM) cells.Methods In vitro, RM cells were cultured and randomized into normal and oxygen-glucose deprivation/reperfusion (OGD/R) groups, and the expression lev-el of circHIPK3 in each group was detected by RT-qPCR.The circHIPK3 lentiviral vector with puromycin resist-ance was constructed, and the overexpression (OE) group and negative control (NC) group were set up.The opti-mal multiplicity of infection (MOI) for RM cells was determined based on fluorescence expression, and puromycin was used to screen RM cells stably expressing circHIPK3 .The cells of OE and NC groups were treated with OGD/R, and the expression levels of ionized calcium binding adaptor molecule 1 (Iba-1) and eukaryotic tumor necrosis factor receptor superfamily (CD40) were detected by Western blot.The circHIPK3 translational protein potential was analyzed by the circRNAdb database, while the potential binding microRNAs on circHIPK3 were predicted by circBank and Starbase databases.Results OGD/R down-regulated circHIPK3 in RM cells (P <0.0001).The sequencing results were accurate and the lentiviral vector of circHIPK3 was constructed successfully.The optimal MOI of RM cells was 80 , puromycin at a concentration of 2μg/ml was used to screen RM cell lines stably express-ing circHIPK3 .RT-qPCR results showed that the expression level of circHIPK3 was significantly higher in the OE group compared with the NC group (P<0.01) .Western blot results revealed that the expression levels of Iba-1 and CD40 in the OE group were markedly lower than those in the NC group (P<0.05) .Protein translation analy-sis showed that circHIPK3 encoded a polypeptide of 404 amino acids with two internal ribosome entry sites (IRES) and an open reading frame (ORF) .Database analysis uncovered that circHIPK3 could target eight specific miR-Nas, namely hsa-miR-3529-5p, hsa-miR-379-5p, hsa-miR-506-3p, hsa-miR-33, hsa-miR-450b-5p, hsa-miR-551b-3p, hsa-miR-193, and hsa-miR-508-3p.Conclusion The overexpression of circHIPK3 effectively suppres-ses OGD/R-induced activation of RM cells.It has the potential to encode peptides and may act as a miRNA sponge.These findings provide a foundation for further study of circHIPK3 functions.

Objective:Based on the business reengineering theory, to construct the emergency nursing process of acute massive hemoptysis in hospital and explore the effect of it.Methods:According to the order of hospitalization, 200 patients with acute massive hemoptysis admitted to the emergency department in Shanghai Pulmonary Hospital Affilated to Tongji University from January to June in 2020 were selected as the control group, and the original treatment process was adopted. Two hundred patients admitted to the emergency department of this hospital from July to December in 2020 were selected as the experimental group, and the reconstructed in-hospital emergency nursing process for acute massive hemoptysis was applied. In both groups, the researchers recorded the time spent in each sub-process by using the in-hospital treatment time record of acute massive hemoptysis and collected the clinical outcomes of patients through electronic cases. The time-consuming, treatment efficiency and patient outcome of each link of the treatment process in the backyard of the two groups were compared.Results:Finally, 192 patients were enrolled in the control group and the experimental group. The median time from admission to intravenous use of hemostatic drugs and from admission to endovascular treatment in the experimental group were 21.50(20.00, 22.50) and 82.50(79.50, 84.50) min, which were lower than those in the control group which spent 40.87(37.06, 44.43) and 135.50(123.50, 147.00) min, and the differences between the two groups were statistically significant ( Z=-16.84, 16.63, both P<0.01). The incidence of asphyxia caused by acute massive hemoptysis in the experimental group was 4.2%(8/192), which was lower than that in the control group, which was 13.0%(25/192) . There was a statistically significant difference in the incidence of asphyxia between the two groups( χ2=9.58, P<0.01). Conclusions:The emergency nursing process of acute massive hemoptysis effectively shortens the time of in-hospital treatment of acute massive hemoptysis, further promotes the seamless connection of multiple links of in-hospital treatment of acute massive hemoptysis, and improves the overall efficiency of in-hospital treatment of patients with acute massive hemoptysis.

Objective:To summarize the characteristics of genetic variation and prenatal diagnosis in pedigrees with X-linked adrenoleukodystrophy (X-ALD) and elucidate the value of prenatal diagnosis in preventing the birth of children with X-ALD.Methods:Twenty pedigrees, clinically diagnosed with X-ALD in Peking University First Hospital from November 2012 and March 2019, were included in this retrospective study. Genomic DNA was extracted from peripheral blood and amniotic fluid or chorionic villi samples of probands and their families for detecting variants in ATP-binding cassette subfamily D member 1 ( ABCD1) gene using polymerase chain reaction (PCR)-Sanger sequencing. Linkage analysis was also performed on five microsatellite markers near ABCD1 gene to exclude maternal contamination. Characteristics of ABCD1 gene variants and prenatal diagnosis of X-ALD pedigrees were summarized by descriptive statistics. Results:Twenty ABCD1 gene variants were identified in the 20 pedigrees. The variants in three probands that were not detected by next-generation sequencing were identified by PCR-Sanger sequencing. Among the mothers of the 20 probands, 17 carried ABCD1 variants and three did not. We performed 24 prenatal diagnoses on 20 pregnancies (24 fetuses) and identified eight fetuses with variants who were finally terminated. The 16 cases without variants were born alive. The validation results obtained after termination or delivery were consistent with those performed prenatally. Conclusions:No hotspot variants in ABCD1 gene are detected in these X-ALD patients and most variants are maternally inherited. PCR-Sanger sequencing is an effective method for detecting ABCD1 variants. Prenatal diagnosis for mothers who had a body with X-ALD could prevent another one from birth.

Objective:To investigate the cognitive level of wearing and removing personal protective equipment in COVID-19 isolation wards of Wuhan Fangcang shelter hospital, and provide the scientific basis for the training of hospital protection. Methods:Cognitive level of wearing and removing personal protective equipment in Wuhan Fangcang shelter hospital were investigated by using a questionnaire which included 14 related scales, including putting on and off protective equipment, hand hygiene disinfection and so on.Results:The total scale score of non-medical workers was 48.82±9.65, which was lower than the total scale score of doctors (55.46±1.29) and nurses (55.0±2.36), P<0.05. The each score of 14 items in the scale of non-medical workers was all lower than those of doctors and nurses ( P<0.05). There were no significant differences in scale scores among doctors, nurses and medical technicians. The total score of the scale of non-medical workers in later period was 55.23±3.26, which was significantly higher than the total score of 44.50±10.14 in previous period, P<0.001. The total score of nurses and medical technicians in the later period was higher than those in the previous period, and the difference was statistically significant. There was no significant difference between the total score of the scale in the later period and that in the previous period. Conclusions:The cognition level of non-medical workers on wearing and removing personal protective devices was lower than those of doctors and nurses, strengthening the practice could improve cognition.

Objective To investigate the clinicopathologic features of Chinese patients with early gastric cancer (EGC) according to the World Health Organization(WHO) diagnostic criteria,and to explore the risk factors of lymph node metastasis (LNM) in EGC.Methods From 2002 to 2017,at the Second Affiliated Hospital of Nantong University,and from 2014 to 2017,at the People's Hospital of Qidong City,the People's Hospital of Haimen City and the People's Hospital of Rugao City,315 EGC patients with complete clinicopathological data were enrolled.The clinicopathologic features were analyzed including gender,age,tumor location,tumor size,macroscopic type,histological type (WHO),differentiation degree,depth of tumor invasion,Lauren subtype,infiltration pattern,ulceration,lymphoid follicles,lymphovascular invasion and perineural invasion.Chi-square test and Fisher's exact test were performed for univariate analysis and the logistic regression was used for multivariate analysis.Results The ratio of EGC was 10.0％ (315/3 140) of patients with gastric cancer and received surgery during the same period.Among the 315 EGC,the ratio of well-differentiated tubular adenocarcinoma was 11.7％ (37/315),which was lower than 24.8％(2 752/11 104) in Japan and 19.9％(41/206) in South Korea,and the differences were statistically significant (x2 =28.208,P＜0.01;x2 =6.51 0,P =0.011),however there was no statistically significant difference when compared with Western countries (11.9 ％,8/67;x2 =0.002,P=0.964).Among the 276 patients who underwent radical gastrectomy,49 (17.8 ％) patients had with LNM.The results of univariate analysis showed that tumor size,macroscopic type,differentiation degree,depth of invasion,infiltration pattern,ulceration and lymphovascular invasion were related with LNM(x2=9.327,6.038,6.381,34.983,19.309,52.297 and 5.058;all P＜0.05).The results of multivariate analysis revealed that lymphovascular invasion and ulceration were the independent risk factors of LNM (odd ratio (OR)=7.028 and 2.566,both P＜0.05).Conclusions There is obvious difference in pathological diagnostic standard of well-differentiated tubular adenocarcinoma between China,Japan and South Korea,which may influence the therapeutic strategy of EGC.Lymphovascular invasion and ulceration are independent risk factors of LNM in EGC.

The editorial board and publication ethics committee of the CEO decided to retract this paper from our journal.

OBJECTIVES: To investigate the surgical outcomes of different uvulopalatopharyngoplasty (UPPP). METHODS: All subjects underwent overnight polysomnography and were evaluated using the Epworth sleepiness scale (ESS), the Quebec sleep questionnaire and the snoring scale at the baseline and 3 and 12 months following operation. The primary endpoint was the overall effective rate representing the sum of the surgical success rate and effective rate. RESULTS: The overall effective rate at 12 months post surgery was 55.6% for simple UPPP, 95.8% for UPPP+GA, and 92.3% for UPPP+TBA. The surgical success rate at 3 and 12 months postoperation for UPPP+GA or UPPP+TBA was significantly higher than simple UPPP (P<0.05). Marked improvement was observed in all patients in the snoring scale score and the ESS score 3 and 12 months following surgery compared to the baseline (P<0.05 in all). CONCLUSION: UPPP, UPPP+GA, and UPPP+TBA are all effective in improving the surgical outcome of obstructive sleep apnea hypopnea syndrome (OSAHS) patients with multilevel obstruction. UPPP+TBA appears to be the most effective in treating OSAHS patients.
Humans ; Polysomnography ; Prospective Studies* ; Quebec ; Sleep Apnea Syndromes ; Sleep Apnea, Obstructive* ; Snoring ; Tongue* ; Surveys and Questionnaires

Objective To explore molecular mechanisms underlying the in vitro counteracting effect of curcumin on malignant melanoma.Methods Cultured A375 and C8161 human melanoma cells were cultivated in vitro,and randomly divided into several test groups and a control group to be treated with different concentrations of curcumin and dimethyl sulfoxide respectively for different durations.Then,methyl thiazolyl tetrazolium (MTT) assay,Transwell assay,flow cytometry and Western blot were performed to evaluate the effect of curcumin on the proliferation,invasion and cell cycle of,as well as expressions of AKT/mTOR signaling pathway-related proteins in A375 and C8161 cells respectively.Statistical analysis was carried out by using t test.Results MTT assay showed that the treatment with curcumin of 5-35 mg/L for 24-96 hours significantly inhibited the proliferation of both A375 and C8161 cells compared with that with dimethyl sulfoxide (all P ＜ 0.001),and the inhibitory effect was in a dose-dependent manner within the range of 5-15 mg/L for A375 cells and within the range of 5-10 mg/L for C8161 cells,and in a time-dependent manner from 0 to 48 hours for both cells.After treatment for 24 hours,the 50％ inhibitory concentration (IC50) of curcumin against A375 cells and C 8161 cells was 10 mg/L and 5 mg/L respectively.Transwell assay demonstrated that the invasion of A375 and C8161 cells was significantly suppressed by 72-hour treatment with curcumin at 10 mg/L and 5 mg/L respectively (both P ＜ 0.001).Flow cytometry showed that the cell cycle of A375 and C8161 cells was arrested at G2/M phase after 24-hour treatment with curcumin at 10 mg/L and 5 mg/L respectively,with significant differences in the proportion of A375 cells and C8161 cells in G2/M phase between the test group and control group (A375 cells:35.00％ ± 3.54％ vs.120.80％ ± 7.46％,P＜ 0.001;C8161 cells:19.33％ ± 4.04％ vs.85.00％ ± 9.53％,P ＜ 0.001).Western blot revealed that the expressions of AKT/mTOR signaling pathway-related proteins were decreased in A375 and C8161 cells after 24-hour treatment with 10 mg/L and 5 mg/L curcumin respectively.Conclusion Curcumin can inhibit the proliferation and invasion of A375 and C8161 cells,likely by blocking cell cycle and inhibiting activation of the AKT/mTOR signaling pathway.

Objective To study the influence and mechanism of hepatocyte growth factor (HGF) on myotube phenotype by myotube transdifferentiation induced by transforming growth factor-β1 (TGF-β1). Methods C2C12 cells were cultured in differentiation medium to induce myotubes formation. The cells were randomly devided into 3 groups. The control group without growth factor interruption. The induction group was supplemented with TGF-β1 (5 ng/mL) while the inhibition group was supplenmented with both TGF-β1 (5 ng/mL) and HGF (30 ng/mL). After 12 hours, the expressions of connective tissue growth factor (CTGF) protein in myotubes were detected by Western blot, the levels of CTGF mRNA were measured by RT-PCR. Results Compared to the control group, the protein and mRNA levels of CTGF significantly increased in TGF-β1 treated group , whereas the protein and mRNA levels of CTGF were significantly lower in inhibition group than those in induction group (P < 0.05). Conclusion HGF can inhibit the effect of TGF-β1 on the expression of CTGF in myotubes , which provides the evidences on the study of skeletal muscle cell transdifferentiation.

OBJECTIVE: To investigate the expression of myosin heavy chain (MHC) in normal laryngeal muscle and the difference between the adductor and abductor. METHOD: Seven patients with total laryngectomy were enrolled in this study. The adductor muscles were acquired from the lateral cricoarytenoid (LCA) muscle and the abductor muscles were acquired from the posterior cricoarytenoid (PCA) muscle. The expression of myosin heavy chain were detected with fluorescent quantitative reverse transcription polymerase chain reaction (RT-PCR) and immunofluorescence staining respectively. RESULT: (1) MHC-II b was expressed in laryngeal muscles at mRNA levels, and not expressed at the protein level; (2) At both mRNA level and protein level, the expression of MHC-I was higher in the PCA muscles than in the LCA muscles while MHC-II level was higher in the LCA muscles than in the PCA muscles. CONCLUSION (1) MHC-II b protein was not expressed in human laryngeal muscles; (2) Phenotypic differences were significant in laryngeal adductor and abductor muscles based on their different functions. PCA contained larger percentage of MHC-I fibers, while LCA contained more MHC-II fibers.
Aged ; Female ; Humans ; Laryngeal Muscles ; chemistry ; metabolism ; Male ; Middle Aged ; Myosin Heavy Chains ; genetics ; metabolism ; Phenotype ; Protein Isoforms