ObjectiveTo explore the potential mechanism of Dingchuan Granule （定喘颗粒， DG） in the treatment of respiratory syncytial virus （RSV） pneumonia. MethodsA total of 60 male Sprague Dawley （SD） rats were randomly divided into control group， model group， ribavirin group， DG low-dose group， DG middle-dose group， and DG high-dose group， with 10 rats in each group. Except for the control group， rats were administrated with RSV via intranasal drip. After model establishment， the DG low-， middle-， and high-dose groups were administrated via oral gavage with DG at 3.47， 6.93， and 13.86 g/（kg·d） respectively， while the ribavirin group was administrated via oral gavage with ribavirin at 15.75 mg/（kg·d）. The drug was given once daily for one week. The rats in the control group and the model group were not given any drug， only subjected to the grasping action. Twenty-four hours after the last administration， the pathological changes of lung tissues were observed and scored using HE staining. The levels of serum inflammatory factors， including tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， and interleukin-6 （IL-6）， were detected by colorimetry. The protein levels of nuclear factor （erythroid derived 2）-like 2 （Nrf2）， Kelch-like ECH-associated protein 1 （Keap1）， heme oxygenase 1 （HO-1）， and NAD（P）H quinone dehydrogenase 1 （NQO1） in lung tissues were measured by Western Blot. The RSV load as well as the gene expression levels of Nrf2， Keap1， HO-1， and NQO1 in lung tissues were determined by qRT-PCR. The level of reactive oxygen species （ROS） in rat lung tissues was detected using chemiluminescence. The levels of glutathione （GSH） and malondialdehyde （MDA） in rat lung tissues were measured by a microassay. ResultsCompared with the control group， other groups had significant increases in pathological score of lung tissue， RSV load， levels of ROS， MDA， serum TNF-α， IL-1β， and IL-6； decrease in GSH level， increases in expression level of Keap1 protein and its mRNA in lung tissue， and significant decrease in levels of Nrf2， HO-1， expression level of NQO1 protein and its mRNA （P＜0.05）. Compared with the model group， all the above-mentioned indicators in the DG low-， middle-， and high-dose groups and the ribavirin group were improved to varying degree （P＜0.05）. The levels of serum TNF-α， IL-1β， and IL-6 in rats of DG dose groups showed a dose-dependent pattern， the DG high-dose group exhibiting the best effect （P＜0.05）. The DG high-dose group was superior to the DG low- and middle-dose groups in reducing the levels of ROS and MDA， and increasing the level of GSH in lung tissues （P＜0.05）. The DG high-dose group and the ribavirin group had better effect than the DG middle-dose group in reducing the RSV load （P＜0.05）. The DG high-dose group was superior to the ribavirin group in improving the protein levels of Nrf2， Keap1， HO-1， and NQO1 （P＜0.05）. ConclusionDG could inhibit oxidative stress by regulating the Nrf2/Keap1/HO-1/NQO1 signaling pathway to improve pulmonary inflammation and treat RSV pneumonia， with the DG high-dose group showing the best effect.

OBJECTIVES: To explore the molecular mechanism by which lncRNA SNHG15 regulates proliferation, invasion and migration of lung adenocarcinoma cells. METHODS: The lncRNA microarray chip dataset GSE196584 and LncBase were used to predict the lncRNAs that interact with miR-30b-3p, and their association with patient prognosis were investigated using online databases, after which lncRNA nucleolar RNA host gene 15 (SNHG15) was selected for further analysis. The subcellular localization of lncRNA SNHG15 and its expression levels in normal human lung epithelial cells and lung adenocarcinoma cell lines were detected using fluorescence in situ hybridization and qRT-PCR. In cultured A549 cells, the changes in cell proliferation, migration, and invasion following transfection with a SNHG15 knockdown plasmid (sh-SNHG15), a miR-30b-3p inhibitor, or their co-transfection were assessed with EdU, wound healing, and Transwell assays. Bioinformatics analyses were used to predict the regulatory relationship between lncRNA SNHG15 and COX6B1, and the results were verified using Western blotting and rescue experiments in A549 cells transfected with sh-SNHG15, a COX6B1-overexpressing plasmid, or both. RESULTS: LncRNA SNHG15 was shown to target miR-30b-3p, and the former was highly expressed in lung adenocarcinoma, and associated with a poor patient prognosis. LncRNA SNHG15 was localized in the cytoplasm and expressed at higher levels in A549 and NCI-H1299 cells than in BEAS-2B cells. In A549 cells, lncRNA SNHG15 knockdown significantly inhibited cell migration, invasion and proliferation, and these changes were reversed by miR-30b-3p inhibitor. A regulatory relationship was found between lncRNA SNHG15 and COX6B1, and their expression levels were positively correlated (r=0.128, P=0.003). MiR-30b-3p knockdown obviously decreased COX6B1 expression in A549 cells, and COX6B1 overexpression rescued the cells from the inhibitory effects of lncRNA-SNHG15 knockdown. CONCLUSIONS LncRNA SNHG15 may compete with COX6B1 to bind miR-30b-3p through a ceRNA mechanism to affect proliferation, migration, and invasion of lung adenocarcinoma cells.
Humans ; MicroRNAs/metabolism* ; RNA, Long Noncoding/genetics* ; Cell Proliferation ; Cell Movement ; Lung Neoplasms/genetics* ; Adenocarcinoma of Lung ; Neoplasm Invasiveness ; A549 Cells ; Adenocarcinoma/genetics* ; Gene Expression Regulation, Neoplastic ; Cell Line, Tumor

Objective To explore the construction idea and application method of knowledge base and knowledge mining system for the prevention and treatment of diseases in traditional Chinese medicine.Methods Guided by the theory of traditional Chinese medicine(TCM),firstly,the knowledge system of TCM Prevention and treatment was sorted out,and the structure and relationship of TCM Prevention and treatment knowledge base were designed according to the classification method of TCM Prevention and treatment;Secondly,according to the needs of pre treatment research,the ancient and modern literature data sources and knowledge collection methods of TCM pre treatment database are proposed;Then,under the framework of the pre treatment classification system,the core knowledge is studied in the aspects of professional annotation,relationship extraction,knowledge audit,and a variety of data mining algorithms are introduced to analyze and mine the knowledge;Finally,the massive data obtained are combined with big data analysis and computer machine learning to realize intelligent information collection,disease analysis and diagnosis and treatment suggestions.Results Under the guidance of traditional Chinese medicine theory,the knowledge base of traditional Chinese medicine for prevention and treatment of diseases can digitize,digitize and intellectualize the basic knowledge and clinical knowledge of traditional Chinese medicine for prevention and treatment of diseases,and can objectively mine and analyze the data,providing a basis for the service and sharing of knowledge of traditional Chinese medicine for prevention and treatment of diseases.Conclusion The knowledge base of TCM Prevention and treatment is an important way for the digital storage of TCM Prevention and treatment knowledge,and provides literature knowledge support and objective evidence of data mining for TCM Prevention and treatment research.

Objective:To determine the incidence of adrenal incidentalomas(AIs) in patients with diabetes mellitus and the metabolism profiles.Methods:A total of 615 hospitalized patients with diabetes mellitus in the Department of Endocrinology and Metabolism of Peking University People′s Hospital from March 2020 to May 2021 were retrospectively included in this study. AIs were screened by unenhanced chest computed tomography(CT) retrospectively and subsequently confirmed by multiplanar reconstruction. Participants′ physical indicators, metabolic profiles, and adrenal function parameters were collected. Unpaired t test, Mann-Whitney U test, and Chi-Square test were adopted to compare the metabolism profiles between diabetes mellitus patients with or without AIs. Regression models were used to estimate the correlations between AIs and the metabolism profiles such as blood glucose, blood lipids, blood pressure, and the adrenal function parameters.Results:Twenty-seven out of 615 participants were detected with AIs(4.4%). Patients with AIs had higher body mass index, waist circumference, and hip circumference than patients without AIs [(29.4±5.1)kg/m 2vs(26.8±3.8)kg/m 2,P=0.018; (102.3±11.7)cm vs(95.8±10.3)cm, P=0.002; (107.3±10.1)cm vs(101.4±7.6)cm, P=0.008]. The levels of serum uric acid and urinary albumin/creatinine ratio were also significantly increased in patients with AIs [(409.6±118.1)μmol/L vs(357.4±100.6)μmol/L, P=0.009; 21.25(7.49, 180.24)mg/g vs 8.60(4.71, 34.56)mg/g, P=0.010]. Besides, individuals with AIs were also associated with a higher risk of co-existing hypertension( P=0.045). Conclusion:The incidence of AIs in patients with diabetes is 4.4%. The presence of AIs in patients with diabetes may associated with increased risk of obesity and hypertension.

Objective:To establish a rapid method to detect the minimum inhibitory concentration (MIC) of imipenem in Klebsiella pneumoniae carbapenemase-producing Klebsiella pneumoniae (KPC-Kp) based on ompK36 gene′s GD mutation. Methods:This was a methodological evaluation study. A total of 258 isolates of Klebsiella pneumoniae were collected from Lishui Municipal Central Hospital from March 2011 to December 2019. Porin gene ompK36 and carbapenemase genes blaKPC, blaNDM, blaIMP and blaOXA-48 were amplified by PCR and confirmed by sequencing. The MIC was detected and confirmed by microbroth dilution susceptibility test, and the corresponding patterns of genotype and MIC were constructed. Based on the patterns, a method for rapid detection of imipenem MIC by real-time fluorescence PCR (RT-PCR) was designed and established. The 159 isolates of non-repetitive Klebsiella pneumoniae collected by Lishui Disease Prevention and Control Center (CDC) from 2017 to 2019 were used for further verification. The sensitivity and specificity were calculated by fourfold table. Kappa test was used to compare the consistency between RT-PCR and microbroth dilution susceptibility test. Results:Among 258 isolates, 109 isolates did not carry carbapenemase gene, 65 isolates carried ompK36 gene GD mutation, 127 isolates carried blaKPC, 15 isolates carried blaNDM, 9 isolates carried blaIMP, and blaOXA-48 was not detected. With mircobroth dilution susceptibility test as the standard, there were 3 corresponding patterns between the drug resistance gene and the imipenem MIC of Kp: when all the 4 carbapenemase genes were negative, MIC≤1 mg/L, the sensitivity was 100% (107/107) and the specificity was 98.4% (125/127); when blaKPC was positive and ompK36 gene GD mutation was negative, 4 mg/L≤MIC≤16 mg/L, the sensitivity was 88.2% (60/68) and the specificity was 98.8% (164/166); when blaKPC and ompK36 gene GD mutation were both positive, MIC≥32 mg/L, the sensitivity was 96.6% (57/59) and the specificity was 96.6% (169/175). RT-PCR detected blaKPC, blaNDM, blaIMP, blaOXA-48 genes accurately.The RT-PCR results of ompK36 gene GD mutation in the KPC-producing isolates were 100% consistent with the sequencing results. In the 159 isolates from Lishui CDC, the sensitivity and specificity of imipenem MIC detected by RT-PCR were higher than 95% in all 3 patterns with mircobroth dilution susceptibility test as the standard, and Kappa value was 0.971. Conclusion:The RT-PCR based on ompK36 gene GD mutation was helpful to quickly determine the MIC range of imipenem in KPC-Kp.

Objective To analyze the value of serum C1q tumor necrosis factor related protein 12(CTRP12)in early pregnancy in predicting gestational diabetes(GDM).Methods 182 pregnant women who underwent prenatal examinations in our department from March 2020 to October 2022 were selected,serum CTRP12 levels were measured at 6～14 weeks of pregnancy.At 24～28 weeks of pregnancy,they were divided into normal glucose tolerance(n=152,NGT)group and GDM group(n=30,GDM)based on OGTT results.The serum CTRP12 levels of the two groups were compared and its predictive value for GDM was analyzed.Results The incidence rate of GDM was 16.48%.Compared with NGT group,the age,pre pregnancy BMI,FPG and family history of diabetes in GDM group were significantly higher,and the serum level of CTRP12 was significantly lower(P<0.05).BMI before pregnancy,family history of diabetes,FPG and serum CTRP12 were the influencing factors of GDM.Receiver operating characteristic analysis showed that the AUC of serum CTRP12 for predicting GDM was 0.822(95%CI 0.759～0.885),the optimal cut-off value was 5.66 ng/ml,with the sensitivity 68.2%and the specificity 90.3%,respectively.Compared with the high level group of CTRP12,the incidence rate of GDM in the low level group of CTRP12 was significantly higher(P<0.05).Conclusions The serum CTRP12 level in GDM patients decreases in the early pregnancy.CTRP12 as an influencing factor may have certain predictive value for GDM.

Familial glucocorticoid deficiency type 4(FGD4) is a rare autosomal recessive disorder caused by mutations in the nicotinamide nucleotide transhydrogenase(NNT) gene. The article presented clinical data, laboratory results, and genetic mutation findings of a child with FGD4. Additionally, a retrospective analysis of the clinical data of FGD4 patients reported domestically and internationally was conducted, summarizing the types of gene mutations and clinical characteristics. This case identifies novel mutation sites in the NNT gene, providing a basis for the early diagnosis and treatment of FGD4 patients.

Objective:To investigate the infection status of the hepatitis B virus among the population over 18 years old in the Chaoyang District of Beijing from August to December 2020.Methods:A multistage stratified cluster random sampling was carried out on residents over 18 years old who lived in Chaoyang for more than six months in 2020. The estimated sample size was 8 525. A blood sample was collected and tested for HBsAg, anti-HBs, and anti-HBc using the Abbott Microparticle Enzyme Immunoassay method. A questionnaire survey was also conducted.Results:A total of 9 875 residents were included; the overall positive rate of HBsAg, anti-HBc, and anti-HBs was 2.8% (95% CI:2.5%-3.1%), 33.9% (95% CI:33.0%-34.9%) and 45.6% (95% CI:44.6%-46.6%), respectively. The negative rate for all three serological markers was 45.9% (95% CI:44.9%-46.9%). The reported hepatitis B vaccination rate was 22.4%. The positive rate of HBsAg and anti-HBc were lower in group aged 18-28 years (1.9% and 6.8%, respectively) than the others, and that of the male was higher (3.6% and 35.4%, respectively) than the female (2.2% and 32.8%, respectively). The negative rate for all three serological markers increased with age. However, the positive rate of anti-HBs decreased with age. Conclusions:In Chaoyang District of Bejing, HBV prevalence among the population over 18 years old has decreased. The publicity and education about hepatitis B prevention and control should be strengthened to reduce the immunization gap of the population further.

Objective To investigate the mechanism of action of Dingchuan granules in intervening respiratory syncytial virus pneumonia based on network pharmacology and animal experiments.Methods The potential active ingredients and targets of each traditional Chinese medicine in Dingchuan granules were obtained from TCMSP and TCMID databases,and the active ingredients of the drugs in Dingchuan granules were screened according to ADME pharmacokinetic parameters;the potential disease targets of respiratory syncytial virus pneumonia were obtained from Genecards,OMIM,and DisGeNet databases;the protein-interaction networks of intersecting targets were visualized by using String platform;the key core targets were visualized by using David database;and the intersection targets were visualized by using David database.Interaction networks were constructed using the String platform to visualize the intersecting targets;GO functional enrichment and KEGG pathway enrichment analyses of the key core targets were performed using the David database;and then the relevant networks for the intervention of Respiratory Syncytial Virus Pneumonia in the Dingchuan particles were constructed using the Cytoscape software(3.9.1).Respiratory syncytial virus(RSV)-induced respiratory syncytial virus pneumonia in young rats was selected for experimental validation.Results The results of the network pharmacology showed that the 177 potential active ingredients of Dingchuan granules acted on 144 targets,and there were 1075 targets related to respiratory syncytial virus pneumonia,and 55 drug-disease intersecting targets were obtained,of which 25 core targets were ALB,IL6,CASP3,EGFR,VEGFA,etc.The GO function of Dingchuan granules was also investigated,and the GO function of Dingchuan was investigated.The results of GO function enrichment analysis showed that the biological processes involved mainly include positive regulation of transcription,positive regulation of RNA polymerase II promoter transcription,positive regulation of gene expression,negative regulation of apoptosis,etc.The KEGG pathway enrichment mainly involves the PI3K-Akt signaling pathway,the cancer pathway,the tumor necrosis factor signaling pathway,the IL-17 signaling pathway and so on.The KEGG pathway enrichment mainly involves PI3K-Akt signaling pathway,tumor necrosis factor signaling pathway,IL-17 signaling pathway,etc.Animal experiments initially showed that the fixed wheezing granules can play a role in inflammation and immune response by regulating the PI3K-Akt signaling pathway,thus participating in the inflammatory and immune response.Compared with the normal group,the ratios of p-PI3K/PI3K and p-Akt/Akt in the lung tissues of young rats in the model group were significantly higher(P<0.05),the viral load was significantly higher(P<0.05),the pathological score of lung tissue damage was significantly higher(P<0.05),and the content of inflammatory factors IL-6 and TNF-α in alveolar lavage fluid(BALF)was significantly higher(P<0.05).Compared with the model group,the expression of p-PI3K/PI3K and p-Akt/Akt proteins in the lung tissues of young rats in each dose group and the positive control group was reduced(P<0.05),the viral load was significantly reduced(P<0.05),the pathological scores of lung tissue injury were significantly reduced(P<0.05),and the contents of inflammatory factors IL-6 and TNF-α in BALF were significantly reduced(P<0.05).Conclusion The study revealed the synergistic mechanism of multi-component,multi-target,and multi-pathway action of Dingchuan granules for the treatment of respiratory syncytial virus pneumonia.It was verified by animal experiments that RSV infection in young rats probably activated the PI3K-Akt signaling pathway,which caused the release of inflammatory factors IL-6 and TNF-α.Dingchuan granules could effectively down-regulate the PI3K-Akt signaling pathway,inhibit the release of inflammatory factors IL-6 and TNF-α,and thus achieve the anti-inflammatory effect.

Objective To analyze the individualized CTV-to-PTV margin dose and positioning errors in radiotherapy for esophageal cancer for improving the treatment accuracy while meeting dose requirements.Methods Fifty-four esophageal cancer patients admitted to Sun Yat-sen University Cancer Center at Huangpu District from June 2021 to June 2022 were enrolled.All of the patients underwent CBCT scans in each fraction,and a total of 1283 CBCT images were collected.The image registration between CBCT image before radiotherapy and planning CT image was carried out to obtain errors in vertical(VRT),longitudinal(LNG),lateral(LAT),Roll,Pitch,and YAW directions.The mean values of six-dimensional positioning errors in the first 5 fractions were calculated,and the results were compared with the total fractional errors using the single sample t-test method for determining the differences.The CTV-to-PTV margin was calculated with the formula(margin=2.5∑+0.7δ),and the calculated margins were divided into 5 groups:Group A(5 mm expansion in all directions),Group B(7.9 mm expansion in LAT direction,and 5 mm expansion in other directions),Group C(11.03 mm expansion in LNG direction,and 5 mm expansion in the other directions),Group D(6.36 mm expansion in VRT direction,and 5 mm expansion in the other directions),and Group E(7.9 mm expansion in LAT direction,11.03 mm expansion in LNG direction,and 6.36 mm expansion in VRT direction).Simulation planning was conducted for 10 patients.Results The proportions of differences between the mean values of six-dimensional errors in the first 5 fractions and the total fractional errors in 54 patients were analyzed.There was no significant difference in 192 out of the 324 directions in 54 patients,accounting for 59.26%(P>0.05).Among them,the LAT,LNG,VRT,Pitch,Roll and YAW directions accounted for 64.81%,57.41%,51.85%,64.81%,57.41%and 59.26%of the total cases.The calculated CTV-to-PTV margin was 7.90,11.03 and 6.36 mm in LAT,LNG and VRT directions.The statistical analysis showed that the differences in the coverage rates of organs-at-risk and target areas among the 5 groups of CTV-to-PTV margins were trivial(P>0.05).Conclusion Using the positioning errors in the first 5 fractions of radiotherapy for esophageal cancer to predict subsequent positioning errors is feasible.The reasonable individualized margin in radiotherapy for esophageal cancer can reduce the inter-fractional off-target rate without increasing the dose delivered to organs-at-risk.The study provides a reference for the target volume margin of esophageal cancer and an important basis for precision treatment.