Animal experiments are essential tools in biomedical research, serving as a bridge between basic research and clinical trials. Systematic reviews and meta-analyses (SRs/MAs) of animal experiments are crucial methods for integrating evidence from animal experiment, which can facilitate the translation of findings into clinical research, reduce translational risks, and promote resource integration in basic research. With the continuous development of the Grading of Recommendations, Assessment, Development, and Evaluation (GRADE) methodology, its application in SRs/MAs of animal experiments has gained increasing attention. This article first outlines the principles and specific applications of the GRADE methodology in SRs/MAs of animal experiments, including qualitative descriptive systematic reviews, meta-analyses, and network meta-analyses. It then deeply analyzes the misuse of the GRADE methodology in practice, including incorrect evidence grading, improper classification of evidence, misapplication in qualitative systematic reviews, inconsistencies between the documentation of the upgrading and downgrading process and results, and inappropriate use for making recommendations. Furthermore, this article comprehensively discusses the factors influencing the grading of evidence certainty in SRs/MAs of animal experiments, including the impact of bias risk, indirectness, inconsistency, imprecision, and publication bias on evidence downgrading, as well as the role of large effect sizes and cross-species consistency in evidence upgrading. Finally, in response to the issues discussed, improvement strategies are proposed, including further research and optimization of the GRADE methodology for SRs/MAs of animal experiments, the development of reporting guidelines tailored to the characteristics of SRs/MAs in animal experiment research, and enhanced professional training for researchers in the GRADE methodology. This article aims to improve the quality of evidence in SRs/MAs of animal experiments, strengthen their reliability in clinical decision-making, and promote the more efficient translation of findings from animal experiment research into clinical practice.

Objective: To retrospectively analyze the epidemiological characteristics of supernumerary teeth in patients aged 4-18 years old and the influencing factors on the selection of anesthesia methods, and to provide a reference for the selection of anesthesia plans for children and adolescents with supernumerary teeth. Methods: This study is a retrospective study approved by the Institutional Ethics Committee. Based on clinical electronic medical record system and cone beam CT (CBCT) data, a retrospective analysis was conducted on 2 210 patients 4-18 years of age who underwent supernumerary tooth extraction at the School of Stomatology, Fourth Military Medical University from August 2019 to July 2021. Inclusion criteria: ① Age 4-18 years old; and ② The American Society of Anesthesiologists (ASA) classifies anesthesia into grades I-II; and ③ Have complete oral and anesthesia case records and relevant imaging data. Exclusion criteria: ① Incomplete medical records or unclear imaging data; and ② Patients with ASA grade II or above. Patients’ gender and age, the number of supernumerary teeth, arch, location, orientation, eruption status, reason for appointment, anxiety level, degree of cooperation, anesthesia method, and other relevant information were collected and statistically analyzed. Results: A total of 1 865 eligible patients were included, with an average age of (8.9±3.2) years old. There were more male patients (71.37%, 1 331 cases) than female patients (28.63%, 534 cases) (P < 0.001), with a gender ratio of 2.49:1. The majority of supernumerary teeth were single (75.97%, 1 417/1 865), primarily located in the maxilla (97.2%, 1 812/1 865) and the anterior dental region (94.2%, 1 757/1 865), and in a centric position (77.3%, 1 442/1 865). The majority of patients with erupted supernumerary teeth were active in seeking treatment (97.67%, 335/343). Patients with supernumerary teeth located in the maxilla and mandible bones, as well as in the nasal floor, were mainly referred to the department for diagnosis, accounting for 94.38%(1 361/1 442) and 90.00% (72/80) (χ2=1 363.24, P < 0.001), respectively. Regarding anesthesia methods, the largest proportion of patients received nitrous oxide sedation-assisted local anesthesia or nerve block anesthesia, accounting for 38.07% (710/1 865), followed by local anesthesia, accounting for 35.23% (657/1 865). The proportion of patients receiving midazolam intravenous sedation with local anesthesia or nerve block anesthesia and general anesthesia was relatively small, accounting for 20.86% (389/1 865) and 5.84% (109/1 865). Patients 13-18 years of age with supernumerary teeth in the mandibular and posterior regions and without anxiety had the highest proportion of local anesthesia use (P < 0.001). In contrast, patients who had supernumerary teeth located at the base of the nose (50%), severe anxiety (94.12%), and poor cooperation (98.18%) had the highest proportion of general anesthesia use (P < 0.001). There was no significant difference (P = 0.35) in the incidence of intraoperative and postoperative complications after the extraction of supernumerary teeth. However, the proportion of anesthesia-related complications, such as dizziness, coughing, and respiratory depression, occurring in patients who received general anesthesia was higher than local anesthesia, accounting for 3.81% (P = 0.006). Conclusion There is a gender difference in the incidence of supernumerary teeth in patients 4-18 years of age, with a higher prevalence in males. The majority of supernumerary teeth are single and located in the maxillary anterior region, predominantly in a centric position. Patients whose teeth had erupted were more likely to seek medical treatment voluntarily, while patients with supernumerary teeth located in the maxilla and mandible bones, as well as in the nasal floor were more likely to be referred to the department. Patients with high levels of anxiety, poor cooperation, young age, multiple teeth, and high surgical difficulty were more inclined to choose general anesthesia.

Ameloblastoma is a benign tumor characterized by locally invasive phenotypes,leading to facial bone destruction and a high recurrence rate.However,the mechanisms governing tumor initiation and recurrence are poorly understood.Here,we uncovered cellular landscapes and mechanisms that underlie tumor recurrence in ameloblastoma at single-cell resolution.Our results revealed that ameloblastoma exhibits five tumor subpopulations varying with respect to immune response(IR),bone remodeling(BR),tooth development(TD),epithelial development(ED),and cell cycle(CC)signatures.Of note,we found that CC ameloblastoma cells were endowed with stemness and contributed to tumor recurrence,which was dominated by the EZH2-mediated program.Targeting EZH2 effectively eliminated CC ameloblastoma cells and inhibited tumor growth in ameloblastoma patient-derived organoids.These data described the tumor subpopulation and clarified the identity,function,and regulatory mechanism of CC ameloblastoma cells,providing a potential therapeutic target for ameloblastoma.

Head and neck squamous cell carcinoma(HNSCC)is characterized by high recurrence or distant metastases rate and the prognosis is challenging.There is mounting evidence that tumor-infiltrating B cells(TIL-Bs)have a crucial,synergistic role in tumor control.However,little is known about the role TIL-Bs play in immune microenvironment and the way TIL-Bs affect the outcome of immune checkpoint blockade.Using single-cell RNA sequencing(scRNA-seq)data from the Gene Expression Omnibus(GEO)database,the study identified distinct gene expression patterns in TIL-Bs.HNSCC samples were categorized into TIL-Bs inhibition and TIL-Bs activation groups using unsupervised clustering.This classification was further validated with TCGA HNSCC data,correlating with patient prognosis,immune cell infiltration,and response to immunotherapy.We found that the B cells activation group exhibited a better prognosis,higher immune cell infiltration,and distinct immune checkpoint levels,including elevated PD-L1.A prognostic model was also developed and validated,highlighting four genes as potential biomarkers for predicting survival outcomes in HNSCC patients.Overall,this study provides a foundational approach for B cells-based tumor classification in HNSCC,offering insights into targeted treatment and immunotherapy strategies.

Objective To summarize the current research status and cutting-edge trends of the off-label drug use in China,with a view to providing reference for researchers in this field.Methods CNKI and SinoMed databases were searched to collect research of the off-label drug use in China,and used Microsoft Excel 2021,the R software Bibliometric,and VOSviewer 1.6.18 to visualize the time and trend of publication,province,issuing authors and units,journals,keywords,and topic evolution of the included studies.Results 1 475 papers were included in the research.A total of 2 808 authors from 31 provinces,cities and regions had conducted relevant studies on over-the-counter medication,with an overall increasing trend in the number of publications.Among them,Guangdong province published the most studies related to this field,the Straits Pharmacy Journal and China Pharmacy published the most studies in this field.Proprietary Chinese medicines,antimicrobials,antitumor drugs,and other drugs were the research hotspots.In addition,the patients in pediatrics,outpatient emergency,obstetrics and gynecology,psychiatry and other departments as a special sick population,the clinical use of medication exists in the overspecification situation was also a future research trend.Conclusion At present,research in this field focuses more on OLDU for special populations,special diseases,special drugs,etc.In the future,researchers should conduct evidence-based evaluation of drugs on the basis of more high-quality evidence in order to seek the best evidence for guiding the clinical use of medication.At the same time,drug administration and medical institutions should also develop standardized management policies and systems to promote the rational and safe use of medication in healthcare institutions.

Metabolic syndrome(MS)is a complex syndrome based on metabolic disorders in the human body,and is a risk factor for cardiovascular diseases and even certain tumors,with a complicated etiology and unclear pathogenesis.Helicobacter pylori(Hp)is one of the most common pathogenic bacteria,closely associated with the occurrence and development of various diseases.Currently,there are numerous studies both domestically and internationally on the relationship between Hp and MS and its components.Most studies suggest that there is an association between Hp and MS and Hp influences the occurrence and development of MS through multiple pathways.Eradicating Hp may become a new option for treating MS.Based on recent studies from both domestic and international sources,this paper discusses the association between Hp and MS,analyzes the effects of Hp on obesity,blood glucose,blood lipids,and blood pressure,and aims to provide new ideas for the prevention and treatment of MS.

GI (GIGANTEA) is one of the output key genes for circadian clock in the plant. The JrGI gene was cloned and its expression in different tissues was analyzed to facilitate the functional research of JrGI. RT-PCR (reverse transcription-polymerase chain reaction) was used to clone JrGI gene in present study. This gene was then analyzed by bioinformatics, subcellular localization and gene expression. The coding sequence (CDS) full length of JrGI gene was 3 516 bp, encoding 1 171 amino acids with a molecular mass of 128.60 kDa and a theoretical isoelectric point of 6.13. It was a hydrophilic protein. Phylogenetic analysis showed that JrGI of 'Xinxin 2' was highly homologous to GI of Populus euphratica. The result of subcellular localization showed that JrGI protein was located in nucleus. The JrGI, JrCO and JrFT genes in female flower buds undifferentiated and early differentiated of 'Xinxin 2' were analyzed by RT-qPCR (real-time quantitative PCR). The results showed that the expression of JrGI, JrCO and JrFT genes were the highest on morphological differentiation, implying the temporal and special regulation of JrGI in the differential process of female flower buds of'Xinxin 2'. In addition, RT-qPCR analysis showed that JrGI gene was expressed in all tissues examined, whereas the expression level in leaves was the highest. It is suggested that JrGI gene plays a key role in the development of walnut leaves.
Juglans/genetics* ; Phylogeny ; Plant Leaves ; Cloning, Molecular ; Gene Expression Regulation, Plant ; Plant Proteins/metabolism*

Objective:To investigate the effects of early Comfort using Analgesics, minimal Sedatives and maximum Humane care (eCASH) patterns on the risk of negative mood and continuous renal replacement therapy (CRRT)-related adverse events in patients with severe CRRT.Methods:A total of 90 patients with severe CRRT in Shenzhen Integrated Traditional Chinese and Western Medicine Hospital from October 2018 to October 2020 were selected as the study subjects, and they were divided into observation group and control group according to random number table method, with 45 patients in each group. The control group was given routine nursing program, and the observation group was given eCASH mode on the basis of the control group.Nursing satisfaction, CRRT-related adverse events and negative mood scores before and after nursing were compared between 2 groups.Results:The nursing satisfaction degree of the observation group was 86.67% (39/45), significantly higher than that of the control group (66.67%, 30/45), and the difference was statistically significant ( χ2=5.03, P<0.05). After nursing, the Hospital Anxiety and Depression Scale-Anxiety(HADS-A) and Hospital Anxiety and Depression Scale-Depression(HADS-D) scores of the observation group were 5.18 ± 0.67 and 5.27 ± 0.61, respectively, lower than 8.14 ± 1.18, 7.94 ± 1.07 before intervention, and 6.33 ± 0.72, 5.94 ± 0.49 of the control group. Barthel Index(BI) (65.17 ± 8.67) was significantly higher than that before nursing 41.56 ± 6.46 and control group 60.48 ± 6.47, the difference was statistically significant ( t values were 5.74-20.76, all P<0.05). The scores of Visual Analogue Scale(VAS), Present Pain Intensity(PPI), sensory total score and emotional total score of observation group after nursing were 3.24 ± 0.56, 1.18 ± 0.25, 6.38 ± 0.89, 2.68 ± 0.59 significantly lower than those before nursing 6.24 ± 0.87, 3.24 ± 0.56, 11.24 ± 1.81, 6.37 ± 1.04 and 4.36 ± 0.67, 1.31 ± 0.31, 7.26 ± 0.96, 2.98 ± 0.62 of the control group. The difference was statistically significant ( t values were 2.19-20.70, P<0.05). Conclusions:eCASH model can significantly improve the negative emotions of patients with severe CRRT, improve their comfort and reduce the risk of related adverse events, which is worthy of clinical promotion.

Objective : To investigate the efficacy of a holistic approach for postoperative pain management in children receiving dental treatment under general anesthesia in day-surgery operating room. Methods: A total of 120 children, aged 3-7 years, of American Society of Anesthesiologists physical status Ⅰ or Ⅱ, with ≥ 10 treated teeth, receiving comprehensive dental treatment under general anesthesia from January 2020 to August 2020 were enrolled in this trial and randomly allocated into the holistic approach group (group H, n=60) and including preemptive analgesia, instructions to parents for pain management and web-based assessment system (assessment pain by scanning the quick response code 4, 6, and 24 hours postoperatively) and the control group (group C, n=60) only scanning the quick response code. Pain, face, legs, activity, cry and consolability (FLACC) scale was used to assess the level of pain 2 h postoperatively and the parents postoperative pain measure (PPPM) was used to assess the level of pain 4, 6, and 24 h postoperatively in two groups. Results: The FLACC scores of group H 2 h postoperatively were significantly lower than group C (P <0.05). The incidences of significant pain (PPPM scores ≥ 6) 4, 6 and 24 h postoperatively in group H were lower than group C (P <0.05). Altogether, 91.7% of parents in group H and 71.6% in group C assessed the level of pain of children over time. The compliance rate of parents in group H was significantly higher than group C (P <0.05). Conclusion The holistic approach had a positive effect on reducing postoperative pain for children receiving dental treatment under general anesthesia in the day-surgery operating room.

Objective:To evaluate the effects of a single general anesthesia on neurodevelopment in preschool children.Methods:American Society of Anesthesiologists physical status Ⅰ-Ⅱchildren of either sex, aged 3-6 yr, who were undergoing dental treatment under general anesthesia or non-general anesthesia, were enrolled in the study.Children in general anesthesia group received dental treatment under general anesthesia, while children in control group received pit and fissure sealant.In general anesthesia group, the children entered the operating room accompanied by their parents and inhaled 50% nitrous oxide and 8% sevoflurane, propofol 2 mg/kg, fentanyl 2 μg/kg, and cisatracurium 0.1 mg/kg and dexamethasone 0.2 mg/kg were intravenously injected, mechanical ventilation was performed after nasotracheal intubation under the guidance of a video laryngoscope, and anesthesia was maintained through inhalation of 2.5%-3.0% sevoflurane.The outcomes of neurodevelopment were assessed and scored using Wechsler Preschool and Primary Scale of Intelligence, Behavior Rating Inventory of Executive Function, and Adaptive Behavior Assessment System.Results:Forty-four children were included in general anesthesia group and 19 children in non-general anesthesia group.There were no significant differences between the two groups in the changes in the scores of Wechsler Preschool and Primary Scale of Intelligence, Behavior Rating Inventory of Executive Function, and Adaptive Behavior Assessment System before anesthesia and at 6 months after operation ( P>0.05). Conclusions:A single general anesthesia exerts no significant effect on neurodevelopment in preschool children.