Apical microsurgery is accurate and minimally invasive, produces few complications, and has a success rate of more than 90%. However, due to the lack of awareness and understanding of apical microsurgery by dental general practitioners and even endodontists, many clinical problems remain to be overcome. The consensus has gathered well-known domestic experts to hold a series of special discussions and reached the consensus. This document specifies the indications, contraindications, preoperative preparations, operational procedures, complication prevention measures, and efficacy evaluation of apical microsurgery and is applicable to dentists who perform apical microsurgery after systematic training.
Microsurgery/standards* ; Humans ; Apicoectomy ; Contraindications, Procedure ; Tooth Apex/diagnostic imaging* ; Postoperative Complications/prevention & control* ; Consensus ; Treatment Outcome

Gene regulation relies on the precise binding of transcription factors (TFs) at regulatory elements, but simultaneously detecting hundreds of TFs on chromatin is challenging. We developed cFOOT-seq, a cytosine deaminase-based TF footprinting assay, for high-resolution, quantitative genome-wide assessment of TF binding in both open and closed chromatin regions, even with small cell numbers. By utilizing the dsDNA deaminase SsdAtox, cFOOT-seq converts accessible cytosines to uracil while preserving genomic integrity, making it compatible with techniques like ATAC-seq for sensitive and cost-effective detection of TF occupancy at the single-molecule and single-cell level. Our approach enables the delineation of TF footprints, quantification of occupancy, and examination of chromatin influences on TF binding. Notably, cFOOT-seq, combined with FootTrack analysis, enables de novo prediction of TF binding sites and tracking of TF occupancy dynamics. We demonstrate its application in capturing cell type-specific TFs, analyzing TF dynamics during reprogramming, and revealing TF dependencies on chromatin remodelers. Overall, cFOOT-seq represents a robust approach for investigating the genome-wide dynamics of TF occupancy and elucidating the cis-regulatory architecture underlying gene regulation.
Transcription Factors/genetics* ; Humans ; Chromatin/genetics* ; Animals ; Binding Sites ; Mice ; DNA Footprinting/methods*

Objective To establish human and mouse pancreatic cancer cell strains stably expressing Cas9 protein,green fluorescent protein,red fluorescent proteins,luciferase-tdTomato,and to validate the activity of luciferase and gene editing of Cas9 function for pancreatic cancer research using luciferase and CRISPR/Cas9 system.Methods In human pancreatic cancer cells(AsPC-1,CFPAC-1,HPAC,BxPC-3,HS 766T,MIA PaCa-2,PANC-1,and SW 1990),and mouse pancreatic cancer cell(Pan02),the cells were infected with Cas9-expressing plas-mid pLv-EF1α-Cas9m1.1-Puro,and single-cell clones were selected for culture and expansion.After extracting the total protein,Western blot verified the expression level of Cas9;Infected with fluorescent protein expression plasmids pLv-EF1α-EGFP,pLv-EF1α-mCherry,pLv-EF1α-tdTomato,pLv-EF1α-Luc2-tdT,and selected single cell clones stably expressing fluorescent proteins were cultured and amplified under fluorescence microscope.Cas9 stable expression cell line was selected to be infected with pLv-EF1α-Luc2-tdT,and the mono-clonal culture of stable expression of fluorescent proteins was selected for expansion under fluorescence micro-scope.One of the cell lines were selected to be infected with Lv-EF1a-mCherry,and the mCherry-positive cells were sorted out by flow cytometry,and then the guide RNA of mCherry gene was then infected by lentivirus to tar-get the mCherry gene,and after cell expansion,mCherry knockdown was detected by fluorescence microscope observation and flow cytometry;5 BALB/c Nude mice were subcutaneously inoculated with MIA PaCa-2-Luc2-tdT cells(1.0×107/cells each),and imaged in vivo after 36 days.Results 48 human pancreatic cancer cell strains with stable Cas9 expression were screened(including 23 cells expressing Cas9m1.1,25 cells expressing Cas9m1.1-Luc2-tdT),33 pancreatic cancer cell strains with stable expression of fluorescent proteins were screened(8 cells expressing EGFP,7 expressing mCherry,and 9 each expressing Luc2-tdT and tdTomato).Cells expressing mCherry and Cas9 were infected with mCherry gRNA and mCherry was knocked down.In vivo imaging showed that both bioluminescence and fluorescence luminescence were present in MIA PaCa-2 cells ex-pressing Luc2-tdT.Conclusions 33 pancreatic cancer cell strains with stable expression of fluorescent proteins are successfully established,in which the Luc2-tdT-expressing cell strains have luciferase activity;48 pancreatic cancer cell strains with stable expression of Cas9 are successfully established,and the Cas9 protein has gene edi-ting activity,gene editing activity varies depending on the original cell strains.

Objective To investigate the efficacy of video-assisted teaching ward rounds for typical cases of oph-thalmology in clinical practice for medical undergraduates.Methods A total of 45 students from clinical medicine in Peking Union Medical College were enrolled.They were randomly divided into the experimental group(23 students)and control group(22 students).Primary angle closure glaucoma(PACG)was selected as the learning content,the experimental group was trained by video-assisted teaching ward rounds,while the control group was trained by internship in outpatient clinic.All students joined the test of typical case PACG and a ques-tionnaire survey was implemented for the experimental group.Results The average score of experimental group was higher than control group[(84.30±4.53)vs.(78.05±5.76),t＝4.05,P＜0.001].For video-assisted teaching ward rounds,in experimental group,86.9%(20/23)students believed it was more practical,95.6%(22/23)students thought it was very helpful and 91.3%(21/23)students considered it was conductive to im-prove interest of learning and clinical thinking ability.Conclusions Video-assisted teaching ward rounds improves students'understanding memory of ophthalmic typical cases,which is a potential supplement teaching method in clinical practice training of ophthalmology for medical undergraduates.

Carcinoma ; Humans ; Larynx

Background::Carbon monoxide (CO) poisoning remains a major cause of accidental injuries and multiple studies have indicated that CO is also associated with significantly severe or long-term toxicity to the central nervous system. Given that CO poisoning causes serious morbidity and mortality, a better understanding of epidemiological features and clinical characteristics of acute CO poisoning in China is crucial.Methods::We collected the clinical data of acute CO poisoning in patients between November 2019 and April 2020 across Shandong province, China and analyzed its characteristics focusing on the weekly amount and the severity of the confirmed cases.Results::A total number of 21,088 acute CO poisoning cases were diagnosed. The overall incidence of acute CO poisoning was approximately 0.021%. On severity rankings, 63% of confirmed cases ( n = 13,378) were mild, 27% (n = 5635) were moderate, and 10% ( n = 2075) were severe. Interestingly, the coastal cities had more confirmed cases than the inland/suburban areas in Shandong. Meanwhile, the number of confirmed cases was negatively correlated with the local mean daily temperature ( P = 0.0167). Conclusions::Mild acute CO poisoning cases accounted for the majority of all confirmed cases during the winter of 2019. In Shandong province, which is located in east China, residents of the coastal cities are more susceptible to CO poisoning than residents of inland cities.

This study was designed to determine the metabolites of Zhali Nusi Prescription(ZLNSP) in rats. The ultra-high performance liquid chromatography-LTQ Orbitrap mass spectrometric(UHPLC-LTQ-Orbitrap-MS) and mass defect filter techniques were applied to analyze the metabolites of ZLNSP in rat plasma, bile, urine and feces. The biological samples were analyzed by ACQUITY UPLC BEH T_3 column(2.1 mm×100 mm,1.7 μm), with 0.1% formic acid water(A)-acetonitrile(B) as mobile phase, and the biological samples were analyzed in negative ion mode by electrospray ionization mass spectrometry(ESI-MS). An analytical method for biological samples of rats was established, and 8 prototype components and 36 metabolites were identified. The results showed that the metabolic pathways of the main components of ZLNSP in rats included methylation, glucuronidation, sulfation and so on. It provi-ded information for the therapeutic effect of ZLNSP in vivo.
Administration, Oral ; Animals ; Bile ; Chromatography, High Pressure Liquid ; Feces ; Plasma ; Prescriptions ; Rats

Objective The objective of this study was to investigate the proliferation,autophagy and the potential mechanism of Ubiquitin-like with PHD and ring finger domains 1(UHRF1)in lung adenocarcinoma cells. Methods The expression of UHRF1 in lung adenocarcinoma tissues was determined by the bioinformatics website(TCGA). The expression of UHRF1 in lung adenocarci-noma cell lines(PC-9,A549 and H1299)and human bronchial epithelial cells(16HBE)was detected by qRT-PCR and Western blot. After transfection of UHRF1-shRNA,CCK-8,clone formation and ki67 were performed to detect the changes in the prolifera-tive capacity of lung adenocarcinoma A549 cells. Western blot was used to detect the changes of autophagy-associated proteins(LC3-I/II and Beclin-1)and proliferation-related proteins(CDK6,Rb and PCNA). Transmission electron microscopy was used to ob-serve the effect of UHRF1 on autophagosomes in A549 cells. Results The expression of UHRF1 in lung adenocarcinoma tissues was significantly higher than that in adjacent tissues. Compared with normal bronchial epithelial 16HBE cells,the mRNA and protein levels of UHRF1 in lung adenocarcinoma A549 and H1299 cells were significantly increased. In addition,CCK-8 assay and colony forma-tion experiments showed that silencing UHRF1 reduced the growth of A549 cells. Ki-67 immunofluorescence staining showed that the proliferation ability of A549 cells after knocking out UHRF1 was significantly lower than that in the normal control group. Further-more,knockdown of UHRF1 resulted in an increased expression of CKD6 and PCNA proteins in comparison with the control-siRNA group. The expression of Rb protein was down-regulated in the UHRF1-siRNA group. Silencing UHRF1 increased the ratio of LC3-II/LC3-I, induced up -regulation of Beclin -1 expression and promoted the formation of autophagic bodies in A549 cells. Conclusion UHRF1 is highly expressed in lung adenocarcinoma,and silencing UHRF1 can inhibit proliferation. This effect may be regulated by promoting autophagy.

In , had used Qimen (LR 14) for the most times compared with other acupoints among the limited numbers of acupuncture prescriptions. Qimen (LR 14) is the front- point of the liver meridian, acting on smoothing liver and regulating activity. It is commonly used to treat the disorders caused by retarded circulation of and blood, such as distension and pain of the hypochondriac region and masses. Based on the opening-closing-pivot theory of the six meridians, the specificity of the running course of the meridian and the property of Qimen (LR 14), the mechanisms of Qimen (LR 14) were analyzed on the treatment of febrile diseases, aiming to enhance the considerations and application of this points in clinical practice and contribute to the further understanding of ' s academic thoughts on the syndrome differentiation in the treatment of diseases.
Acupuncture Points ; Acupuncture Therapy ; Fever ; therapy ; Humans ; Meridians ; Pain

Objective To study antioxidant role and mechanism of Rg1 in rats with cerebral ischemia reperfusion injury (IRI).Methods One hundred and twenty healthy male rats were randomly divided into six groups: control group, sham operation group, model group, different concentration (30,60,90 mg/kg) of Rg1 treatment group.MCAO SD rats model was established by suture－occluded method;the Rg1 treatment groups were given Rg1 i.p. in advance, after 24 hours of reperfusion, neurobehavioral scores of all groups were examined by Longa’s standard;The expression of Nrf2 and HO－1 were analyzed by western blot;The content of SOD and MDA was detected by kit.Results The score of model group rats are significantly higher than control group,compared with the model group, the score of different concentration of Rg1 treatment group was decreased (P＜0.05) . The Nrf2 and HO－1 expression in model group was mildly higher than the control or sham group (P＜0.05) . Both of them in every Rg1 treatment group was higher than model group. Compared with control or sham group, SOD content was observably depressed but MDA content was dramatically increased in model group,interestingly,SOD content was enhanced, MDA content was attenuated in different concentration of Rg1 treatment group (P＜0.05). Conclusion Rg1 increases Nrf2 and HO－1 protein expression and SOD content, reduces MDA content,improves neurofunctional performance of rats after MCAO,and alleviates cerebral cerebral IRI.