ObjectiveTo investigate the potential mechanism of action of the Qufeng Tongqiao Cough-relieving Decoction （祛风通窍止咳方， QTCD） in the treatment of upper airway cough syndrome （UACS）. MethodsTwenty-four guinea pigs were randomly divided into blank group， model group， traditional Chinese medicine （TCM） group， and inhibitor group， with six guinea pigs in each group. Except for the blank group， guinea pigs were sensitized with ovalbumin and aluminum hydroxide via intraperitoneal injection， followed by ovalbumin nasal drops combined with smoke exposure to establish the UACS model. After modeling， the TCM group was administered QTCD 0.9 g/（100 g·d） by gavage， the inhibitor group received the transient receptor potential vanilloid receptor 4 （TRPV4） inhibitor GSK2193874 1 mmol/L， 5 min by nebulisation， and the blank group and model group were given 2 ml/（100 g·d） normal saline by gavage once daily. After 7 days of treatment， a cough provocation test was performed using 0.4 mol/L citric acid. The levels of IgE in serum and inflammatory cytokines， including interleukin-6 （IL-6）， interleukin-8 （IL-8） in serum， bronchoalveolar lavage fluid （BALF）， and nasal lavage fluid （NLF） were detected by enzyme-linked immunosorbent assay （ELISA）. Histopathological changes in lung and nasal mucosal tissues were observed by hematoxylin-eosin （HE） staining. Immunohistochemistry was used to detect the protein levels of TRPV4， substance P （SP）， and calcitonin gene-related peptide （CGRP） in lung and nasal mucosal tissues. Real-time polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression of TRPV4， SP， and CGRP in lung tissues. ResultsHE staining showed significant structural damage and infiltration of inflammatory cells in the lung and nasal mucosal tissues in the model group， while the TCM group and inhibitor group showed improved pathological changes. Compared with the blank group， the model group showed increased cough frequency， serum IgE level， and IL-6 and IL-8 levels in serum， BALF， and NLF. The protein levels of TRPV4， SP， and CGRP in lung and nasal mucosal tissues and their mRNA expression were elevated （P＜0.05 or P＜0.01）. Compared with the model group， the TCM group and inhibitor group showed reduced cough frequency， serum IgE level， and TRPV4 and SP mRNA expression in lung tissues. The TCM group showed reduced IL-6 and IL-8 levels in serum， BALF， and NLF， and reduced TRPV4 and CGRP protein levels in lung and nasal mucosal tissues. The inhibitor group showed reduced IL-6 and IL-8 levels in serum， BALF， and NLF， reduced IL-6 in BALF， reduced IL-8 in NLF， and decreased TRPV4， SP， and CGRP protein levels in lung tissues and SP and CGRP protein levels in nasal mucosal tissues （P＜0.05 or P＜0.01）. Compared with the TCM group， the inhibitor group had increased serum IgE， IL-6， and IL-8 levels， increased IL-6 level in BALF， and increased IL-8 levle in NLF， but decreased SP protein level in lung tissues and increased TRPV4 and SP mRNA expression in lung tissues （P＜0.01）. ConclusionQTCD effectively reduces cough frequency in the UACS guinea pig model. Its mechanism may involve inhibiting the activation of the TRPV4 pathway， improving airway neurogenic inflammation， alleviating inflammatory responses， and reducing cough hypersensitivity.

Objective:To evaluate the effect of neck muscle training on reducing the neck setup error during radiotherapy in patients with nasopharyngeal carcinoma.Methods:Clinical data of nasopharyngeal carcinoma patients who were treated with radiotherapy in the First Affiliated Hospital of Air Force Medical University from February 2021 to October 2022 were selected and analyzed. All subjects were randomly divided into the experimental group ( n=48) and control group ( n=51) at a ratio of 1:1 using a random number table method. In the experimental group, patients received neck muscle training prior to treatment, and those in the control group received conventional treatment without additional interventions. Cone beam computed tomography (CBCT) was performed weekly to measure and analyze the setup errors at the level of the slopes, 4th cervical vertebra (C4) and 7th cervical vertebra (C7). The four-dimensional displacement [left-right (LR), superior-inferior (SI), anterior-posterior (AP), rotation (Rtn)] systematic errors and random errors at each level were calculated, and the planning target volume (PTV) boundary was calculated. The differences at three different levels of slope, C4, C7 (LR, SI, AP axis) were compared between two groups. The correlation of setup errors in each direction was analyzed by Spearman correlation analysis. The changes of cervical curvature, cutaneous toxicity (common terminology criteria for adverse events V3.0) and pain assessment (numerical rating scale) were compared between two groups. Qualitative data between two groups were compared by χ2 test. Quantitative data between two groups were compared by t-test. Results:Baseline features were well balanced in both groups. The setup error in the experimental group was smaller than that in the control group. For the setup error in the AP direction, the setup errors at the levels of slope, C4 and C7 in the experimental group were (0.94±0.88) mm, (1.13±1.03) mm and (1.32±1.22) mm, significantly less than (1.66±1.23) mm, (1.63±1.35) mm and (1.89±1.48) mm in the control group (all P<0.001). In the SI direction, the setup errors at the levels of slope, C4 and C7 in the experimental group were (1.14±0.87) mm, (1.31±0.93) mm and (1.39±0.95) mm, compared with (1.22±0.95) mm, (1.40±1.11) mm and (1.52±1.08) mm in the control group ( P=0.278, 0.272, 0.100). The differences in the AP direction at the level of C4 and C7 in the experimental group were smaller than those in the control group ( P=0.014, 0.005). The required PTV boundary in the AP direction in the experimental group was increased from 1.77 mm at the slope level to 2.98 mm at the level of C7. In the control group, it was increased from 3.02 mm at the slope level to 4.78 mm at the level of C7. Correlation analysis showed that at the C4 and C7 levels, and the setup errors in the SI direction were moderately negatively correlated with those in the AR direction. There were no significant differences in cervical curvature, skin toxicity and pain assessment between two groups. Conclusion:Neck muscle training can reduce the setup error in the AP direction and PTV boundary of radiotherapy in patients with nasopharyngeal carcinoma, which is worthy of clinical promotion.

Chronic alcohol consumption causes liver steatosis, cell death, and inflammation. Melatonin (MLT) is reported to alleviate alcoholic liver disease (ALD)-induced injury. However, its direct regulating targets in hepatocytes are not fully understood. In the current study, a cell-based screening model and a chronic ethanol-fed mice ALD model were used to test the protective mechanisms of MLT. MLT ameliorated ethanol-induced hepatocyte injury in both cell and animal models (optimal doses of 10 μmol/L and 5 mg/kg, respectively), including lowered liver steatosis, cell death, and inflammation. RNA-seq analysis and loss-of-function studies in AML-12 cells revealed that telomerase reverse transcriptase (TERT) was a key downstream effector of MLT. Biophysical assay found that epidermal growth factor receptor (EGFR) on the hepatocyte surface was a direct binding and regulating target of MLT. Liver specific knock-down of Tert or Egfr in the ALD mice model impaired MLT-mediated liver protection, partly through the regulation of nuclear brahma-related gene-1 (BRG1). Long-term administration (90 days) of MLT in healthy mice did not cause evident adverse effect. In conclusion, MLT is an efficacious and safe agent for ALD alleviation. Its direct regulating target in hepatocytes is EGFR and downstream BRG1-TERT axis. MLT might be used as a complimentary agent for alcoholics.

The mechanisms underlying autophagic defects in nonalcoholic steatohepatitis (NASH) remain largely unknown. We aimed to elucidate the roles of hepatic cyclooxygenase 1 (COX1) in autophagy and the pathogenesis of diet-induced steatohepatitis in mice. Human nonalcoholic fatty liver disease (NAFLD) liver samples were used to examine the protein expression of COX1 and the level of autophagy. Cox1^Δhepa mice and their wildtype littermates were generated and fed with 3 different NASH models. We found that hepatic COX1 expression was increased in patients with NASH and diet-induced NASH mice models accompanied by impaired autophagy. COX1 was required for basal autophagy in hepatocytes and liver specific COX1 deletion exacerbated steatohepatitis by inhibiting autophagy. Mechanistically, COX1 directly interacted with WD repeat domain, phosphoinositide interacting 2 (WIPI2), which was crucial for autophagosome maturation. Adeno-associated virus (AAV)-mediated rescue of WIPI2 reversed the impaired autophagic flux and improved NASH phenotypes in Cox1^Δhepa mice, indicating that COX1 deletion-mediated steatohepatitis was partially dependent on WIPI2-mediated autophagy. In conclusion, we demonstrated a novel role of COX1 in hepatic autophagy that protected against NASH by interacting with WIPI2. Targeting the COX1-WIPI2 axis may be a novel therapeutic strategy for NASH.

Ubiquitination is one of the reversible protein post-translational modifications, in which ubiquitin molecules bind to the target protein in a cascade reaction of ubiquitin activating enzymes, ubiquitin conjugating enzymes, and ubiquitin ligases. The deubiquitinating enzymes (DUBs) remove ubiquitin residues from the substrates, which play key roles in the formation of mature ubiquitin, the removal and trimming of ubiquitin chains, as well as the recycling of free ubiquitin chains. Ubp14, a member of the ubiquitin specific proteases family in Saccharomyces cerevisiae, is mainly responsible for the recycling of intracellular free ubiquitin chains. To investigate its global biological function, a ubp14∆ mutant was constructed by homologous recombination technique. The growth rate of ubp14∆ mutant was lower than that of the wild-type (WT) strain. Using stable isotope labeling by amino acids in cell culture (SILAC) combined with deep coverage proteomics analysis, the differentially expressed proteins of ubp14∆ mutant relative to the wild-type strain were systematically analyzed. A total of 3 685 proteins were identified in this study, and 109 differentially expressed proteins were filtered out by statistical analysis. Gene ontology analysis found that differentially expressed proteins caused by Ubp14 loss were mainly involved in amino acid metabolism, REDOX, heat shock stress and etc, which shed light on the broad biological function of this DUB. This study provides highly reliable proteomic data for further exploring the biological functions of the deubiquitination enzyme Ubp14, and further understanding the relationship between the free ubiquitin homeostasis and biological process regulation.
Saccharomyces cerevisiae/metabolism* ; Proteomics ; Endopeptidases/metabolism* ; Ubiquitin/metabolism* ; Ubiquitination ; Proteins/metabolism* ; Deubiquitinating Enzymes/metabolism* ; Biological Phenomena

Objective:To compare the surgical efficacy and cosmetic satisfaction of the transoral endoscopic thyroidectomy by vestibular approach (TOETVA), endoscopic thyroidectomy via bilateral areola approach (BAA), and open thyroidectomy (OT) in the treatment of papillary thyroid carcinoma (PTC) in stage T1b.Methods:A total of 178 patients with PTC in stages pT1bN0M0 and pT1bN1aM0 treated from January 2017 to December 2018 were divided into TOETVA group ( n=59), BAA group ( n=43), and OT group ( n=76). The baseline characteristics, relevant indexes of surgical efficacy and follow-up data were compared between the three groups, and also the surgical efficacy indexes were compared between unilateral lobectomy + central lymph node dissection (unilateral lobectomy subgroup) and bilateral lobectomy + central lymph nodes dissection (total resection subgroup). SPSS 19.0 statistical software was used for data analysis. Results:The mean ages of patients in TOETVA group and BAA group were significantly younger than those in OT group, and the proportion of women in TOETVA group and BAA group was significantly higher than that in OT group, all P<0.05. None of patients in TOETVA and BAA groups were converted to open surgery. Among TOETVA, BAA and OT groups, there were significant differences in the mean operation time [unilateral lobectomy subgroup: (198.0±45.2) min, (162.0±36.9) min and (79.4±28.6) min, P<0.05; total resection subgroup: (230.0±36.0) min, (219.8±68.1) min and (102.8±40.0) min, P<0.05], in total drainage volume [unilateral lobectomy subgroup: (195.0±55.6) ml, (178.1±50.4) ml and (127.0±30.1) ml, P<0.05; total resection subgroup: (221.1±46.7) ml, (245.3±71.2) ml and (137.7±41.6) ml, P<0.05], and the incidence of subcutaneous ecchymosis in TOETVA group or BAA group was higher than that in the OT group [5.1% (3/59), 11.6% (5/43) vs. 0, χ 2=3.952 and 9.225 respectively, both P<0.05]. The mean level of C-reactive protein in TOETVA group was higher than that in OT group [(16.8±10.7) vs. (9.5±6.9), P<0.05]. Following-up in the third month after surgery between three groups in cosmetic satisfaction scores [(2.7±0.5) vs. (2.6±0.7) vs. (1.7±0.8)], scar self-consciousness scores (0[0,1] vs. 1[0,2] vs. 2[1,2]), and quality of life scores [(9.1±1.1) vs. (8.9±1.1) vs. (7.5±0.8)], cosmetic satisfaction and quality of life in TOETVA and BAA group were better than that in OT group, in terms of scar self-consciousness: TOETVA0.05). The median follow-up time of the three groups was 25 months (TOETVA group), 28 months (BAA group), and 32 months (OT group) respectively, without disease progression. Conclusion:TOETVA and BAA are optional surgical methods for PTC in stages pT1bN0M0 and pT1bN1aM0, with good safety and patient's cosmetic satisfaction.

Objective To investigate the role of different parenting styles plays in conduct problems of 3-years-old preschool children.Methods Participants were from 148 preschools in Longhua Shenzhen,China.A structured questionnaire covering family demographic characteristics,Conners' Parent Symptom Questionnaire and Egna Minnen Betraffande Uppfostran-Parents (EMBU-P) were conducted among primary caregivers of each child.There were 9289 valid participants in the analysis and logistic regression was used to assess the relationship between three dimensions of parenting (rejection,emotional warmth and control/overprotection) and conduct problems.Results After allowing for certain confounders,parental rejection (OR =1.17,P＜0.01) and control/overprotection (OR =1.03,P＜0.01) was positively associated with conduct problems and emotional warmth was negatively related to conduct problems (OR=0.98,P＜0.01).Stratified analysis showed a significant relationship between conduct problems and both parents' rejection (OR father=1.16,P＜0.01;OR motber=1.17,P＜0.01),father's emotional warmth (OR=0.96,P＜0.01) and mother's control/overprotection (OR=1.04,P＜0.01).This relationship did not vary between boys and girls and between single and not single children.Conclusion Influence of parenting should be taken into consideration in the prevention and intervention of conduct problems.

[ ABSTRACT] AIM:To investigate the effect of hydrogen sulfide ( H2 S) on airway inflammation induced by ozone (O3) exposure and its mechanisms.METHODS:C57BL/6 mice (n=32) were randomly divided into control group, O3 group, NaHS+O3 group and NaHS group.The mice in O3 group and O3 +NaHS group were exposed to 2.14 mg/m3 O3 for 3 h on days 1, 3 and 5, while the mice in control group and NaHS group were exposed to filtered air .NaHS (14μmol/kg) was administered intraperitoneally to the mice in NaHS group and O 3 +NaHS group 30 min before each exposure .After the last exposure for 24 h, the airway responsiveness was determined , and bronchoalveolar lavage fluid ( BALF) was collected for counting inflammatory cells and measuring total protein concentration .The lung tissues were collected for observing the morphological changes with HE staining .The levels of interleukin-6 ( IL-6 ) , interleukin-8 ( IL-8 ) , malondialdehyde ( MDA) and NF-κB p65 protein in the lungs were determined .RESULTS: Compared with control group , the airway re-sponsiveness, inflammatory cells, protein concentration, inflammation score, levels of IL-6, IL-8, MDA and NF-κB p65 in O3 group increased significantly , but these in NaHS+O3 group decreased compared with O 3 group.CONCLUSION: The present findings suggest that H 2 S attenuates O3 induced airway inflammation by inhibiting NF-κB expression and preventing lipid peroxidation .

Objective To explore the interaction between passive smoking and folic acid supplement during pregnancy on children autism spectrum disorder (ASD) behaviors.Methods Children aged about 3 years were enrolled at kindergarten entrance in Longhua district of Shenzhen in 2014.Self-administered questionnaires were completed by their primary caregivers and the information about children' s age,gender,history of preterm birth and low birth weight,parents' education level,parents' reproductive age and family income were collected.The children ASD behaviors were assessed with Autism Behavior Checklist (ABC).According to the cut point of ABC,the children were divided into normal group with score less than 31,sub-clinical group with score ranging from 31 to 61 and suspect clinical group with score no less than 62.After controlling for potential confounders,multivariate logistic regression analysis was performed to evaluate the main effects and the interaction between passive smoking and folic acid supplement during pregnancy on children ASD behaviors.Results Maternal passive smoking during pregnancy was significantly associated with children ASD behaviors (sub-clinical group:OR=1.48;suspect clinical group:OR=2.85),and maternal folic acid supplement during pregnancy was not related to children ASD behaviors (sub-clinical group:OR=1.04;suspect clinical group:OR=0.75).Stratified analysis showed that folic acid supplement during pregnancy was negatively associated with children ASD behaviors (suspect clinical group:OR=0.30) among children without mothers' passive smoking during pregnancy,and that mothers' passive smoking during pregnancy was positively associated with children ASD behaviors (sub-clinical group:OR=1.52;suspect clinical group:OR=4.45) among the children whose mothers had folic acid supplement during pregnancy.Furthermore,an interaction effect on children ASD behaviors was found between passive smoking and folic acid supplement during pregnancy (suspect clinical group:OR=5.30).Conclusion Passive smoking and folic acid supplement during pregnancy were related to children ASD behaviors and had an interaction on children ASD behaviors.

OBJECTIVETo detect the expression of transient receptor potential canonical 1 (TRPC1) in a mouse model of ozone-induced lung inflammation and explore its role in lung inflammation.

METHODSIn a mouse model of lung inflammation established by ozone exposure, the expression of TRPC1 in the inflammatory lung tissues was detected by RT-PCR, Wstern blotting and immunohistochemistry.

RESULTSCompared to the control mice, the mice exposed to ozone showed significantly increased expression level of TRPC1 mRNA and protein in the inflammatory lung tissues (P<0.05). Immunohistochemistry showed increased TRPC1 protein expressions in the alveolar epithelial cells, bronchial epithelial cells, and inflammatory cells in the inflammatory lung tissues (P<0.05). The mRNA and protein expression levels of TRPC1 were positively correlated with the counts of white blood cells, macrophages, neutrophils and lymphocytes in the bronchoalveolar lavage fluid of the exposed mice (P<0.01).

CONCLUSIONTRPC1 may play a role in ozone-induced lung inflammation in mice.

Animals ; Bronchoalveolar Lavage Fluid ; Disease Models, Animal ; Gene Expression ; Inflammation ; pathology ; Lung ; metabolism ; pathology ; Mice ; Ozone ; adverse effects ; Pneumonia ; metabolism ; pathology ; RNA, Messenger ; TRPC Cation Channels ; metabolism