Objective:To understand the molecular characteristics of the mutant strain of polymerase basic protein 2 (PB2) gene of influenza A (H1N1pdm) in Guangdong province, and to explore its specific molecular sites, so as to provide scientific basis for the prevention and control of influenza virus.Methods:Throat swab samples were collected from 2 cases infected with PB2 gene variant strains for virus isolation, and 23 influenza virus strains were selected from Guangdong province for sequencing analysis. The reference sequences and vaccine strain sequences provided by GISAID were used to perform evolutionary analysis on hemagglutinin (HA) and PB2 genes. Virus strain antigen analysis and neuraminidase (NA) inhibition test were carried out. PB2 protein model was constructed and polymerase activity was analyzed.Results:H399N amino acid mutation occurred in the HA gene of PB2-D701N and PB2-A271S variant strains, both of which belonged to the branch of 6B.1A.5a.2a. They belonged to the same big branch and different small branches as the vaccine strain A/Victoria/4897/2022, and they are all vaccine-like strains. In the three-dimensional structure, the mutations of PB2-D701N and PB2-A271S change charge and hydrophobicity.Conclusions:PB2-D701 and A271 were highly conserved, and PB2 mutant strains were not the dominant strains. The PB2 mutant had high antigenicity with the vaccine. The PB2 mutant strain is sensitive to NA inhibitors. The three-dimensional model predicted that PB2-D701N mutation could enhance virulence and affect transmissibility of influenza virus, while PB2-A271S mutation could affect polymerase activity and polymerase complex synthesis of influenza virus.

BACKGROUND:Ankylosing spondylitis is a chronic inflammatory disease with chronic rheumatic immunity.Soft tissue ossification and fusion and spinal stiffness can cause biomechanical changes. OBJECTIVE:To reconstruct the lumbar-sacral intervertebral disc in ankylosing spondylitis patients with lumbar kyphosis by finite element analysis,and to study the range of motion of each segment of T11-S1 and the biomechanical characteristics of annulus fibrosus and nucleus pulposus. METHODS:The imaging data were obtained from an ankylosing spondylitis patient with lumbar kyphosis.The original CT image data of continuously scanned spine were imported into Mimics 21.0 in DICOM format,and T11-S1 was reconstructed respectively.The established model was imported into 3-Matic software in the format of"Stl"to reconstruct the intervertebral disc,and the fibrous intervertebral disc model was obtained.The improved model was further imported into Hypermesh software,and the vertebra,nucleus pulposus,annulus fibrosus and ligament were mesh-divided.After the material properties were given,the model was imported into ABAQUS software to observe the range of motion of each vertebral body in seven different working conditions of T11-S1,and analyze the biomechanical characteristics of each segment of annulus fibrosus and nucleus pulposus. RESULTS AND CONCLUSION:(1)The range of motion of L1 vertebrae was higher than that of other vertebrae under six different working conditions:extension,forward flexion,rotation(left and right),and lateral flexion(left and right).The maximum range of motion was 2.18° during L1 vertebral flexion,and the minimum range of motion was 0.12° during L5 vertebral extension.(2)The annular fiber flexion at L2-L3 segments was greater than the extension(P<0.05),and the annular fiber flexion at L3-L4 and L4-L5 segments was less than the extension(P<0.05).The left rotation of L1-L2 annular fibers was greater than the right rotation(P<0.05).The left flexion of the annulus was greater than the right flexion in L1-L2,L2-L3,L3-L4,L4-L5 and L5-S1 segments(P<0.05).(3)The nucleus pulposus stresses of T11-L12,L1-L2,L2-L3,L3-L4 and L4-L5 segments in forward flexion were greater than in extension(P<0.05).The left rotation of T12-L1 and L3-L4 segments was smaller than the right rotation(P<0.05),and that of T11-T12,L1-L2,and L2-L3 segments was larger than the right rotation(P<0.05).The left flexion was larger than the right flexion in the T11-S1 segment.(4)It is concluded that in ankylosing spondylitis patients with lumbar kyphosis,the minimum range of motion of the vertebral body is located at the L5 vertebral body in extension.To prevent fractures,it is recommended to avoid exercise in the extension position.During the onset of lumbar kyphosis in patients with ankylosing spondylitis,the maximum stress of the annulus fibrosus and nucleus pulposus is located in the L1-L2 segment,which is fixed and will not alter with the change of body position.The late surgical treatment and correction of deformity should focus on releasing the pressure of the annulus fibrosus and nucleus pulposus in this segment to avoid the rupture of the annulus fibrosus and the injury of the nucleus pulposus.

ObjectiveBased on knowledge mapping， the studies on prediction models in the field of traditional Chinese medicine （TCM） were visually analyzed， which provided a reference basis for the excavation and evolution of the future research direction by combing the development process and summarizing the research hotspots and dynamic trends. MethodChina National Knowledge Infrastructure and Web of Science Core Collection databases were searched to obtain studies on prediction models in the field of TCM from inception to February 28， 2023. Endnote X20 software was used for document management. Knowledge mapping generated by CiteSpace software and VOSviewer software was used to visually analyze the characteristics of publication， institutional cooperation relationship， author cooperation network， co-citation， and keywords. ResultA total of 264 pieces of Chinese literature and 266 pieces of English literature were included， and the overall number of research publications showed an increasing trend year by year. The cooperation relationship between the issuing institutions showed obvious regional characteristics， with the closest cooperation relationship between the universities of TCM and their affiliated hospitals， as well as secondary units subordinate to scientific research institutions. The number of research teams and team members publishing papers in English was higher， and cooperation between different teams was more frequent. Groundbreaking and/or referential studies were widely cited and referred to. The highly cited literature was mainly published in complementary and alternative medicine journals and pharmaceutical journals. Research hotspots mainly focused on clinical prediction models of TCM， quantitative models of TCM， and specific modeling methods. The application of artificial intelligence technologies such as machine learning and deep learning in the field of TCM will be the most cutting-edge research direction in the future. ConclusionThe field of TCM is paying more and more attention to the studies on prediction models， while the research cooperation mode involving multiple organizations and teams has increasingly become the mainstream. With the continuous development of multi-disciplinary integration， studies on prediction models are closely related to the development and rise of innovative techniques and methods， and any breakthrough in theory or application will induce and guide a new round of research upsurge. Systematic reviews of topic-specific prediction models should be carried out in the future to provide evidence-based evidence.

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.

High mobility group box 1 protein（HMGB1）gene is located on chromosome 13q12 and is widely expressed in various tissues and cells.The encoded non-histone chromosome binding protein regulates transcription and participates in DNA repair.It plays an important role in processes such as inflammation，cell differentiation and tumor cell migration，and has become a valuable biomarker in a variety of diseases. In recent years，more and more studies have found that this gene is closely related to various diseases in children，such as respiratory diseases，digestive diseases，autoimmune diseases，cardiovascular diseases and so on.This article reviews the research progress of HMGB1 gene in children ' s diseases.

Cardiovascular System/pathology* ; Constriction, Pathologic/genetics* ; Point Mutation ; Neurofibromin 1/genetics* ; Animals ; Mice

As an essential trace element in living organisms, copper is actively involved in normal physiological processes in various systems and is maintained at low level to achieve copper homeostasis. Copper homeostasis, once being disrupted, would induce cell death, and this new form of cell death is known as copper death. In recent years, copper death has been increasingly recognized as an important factor mediating the onset and progression of central nervous system (CNS) diseases. Therefore, we review the pathogenic mechanism of copper death in CNS diseases, as well as its therapeutic strategies so as to deepen the understanding of researchers.

OBJECTIVE: To establish a rapid detection and genotyping method for SARS-CoV-2 Omicron BA.4/5 variants using CRISPPR-Cas12a gene editing technology. METHODS: We combined reverse transcription-polymerase chain reaction (RT-PCR) and CRISPR gene editing technology and designed a specific CRISPPR RNA (crRNA) with suboptimal protospacer adjacent motifs (PAM) for rapid detection and genotyping of SARS- CoV-2 Omicron BA.4/5 variants. The performance of this RT- PCR/ CRISPPR-Cas12a assay was evaluated using 43 clinical samples of patients infected by wild-type SARS-CoV-2 and the Alpha, Beta, Delta, Omicron BA. 1 and BA. 4/5 variants and 20 SARS- CoV- 2-negative clinical samples infected with 11 respiratory pathogens. With Sanger sequencing method as the gold standard, the specificity, sensitivity, concordance (Kappa) and area under the ROC curve (AUC) of RT-PCR/CRISPPR-Cas12a assay were calculated. RESULTS: This assay was capable of rapid and specific detection of SARS- CoV-2 Omicron BA.4/5 variant within 30 min with the lowest detection limit of 10 copies/μL, and no cross-reaction was observed in SARS-CoV-2-negative clinical samples infected with 11 common respiratory pathogens. The two Omicron BA.4/5 specific crRNAs (crRNA-1 and crRNA-2) allowed the assay to accurately distinguish Omicron BA.4/5 from BA.1 sublineage and other major SARS-CoV-2 variants of concern. For detection of SARS-CoV-2 Omicron BA.4/5 variants, the sensitivity of the established assay using crRNA-1 and crRNA-2 was 97.83% and 100% with specificity of 100% and AUC of 0.998 and 1.000, respectively, and their concordance rate with Sanger sequencing method was 92.83% and 96.41%, respectively. CONCLUSION By combining RT-PCR and CRISPPR-Cas12a gene editing technology, we successfully developed a new method for rapid detection and identification of SARS-CoV-2 Omicron BA.4/5 variants with a high sensitivity, specificity and reproducibility, which allows rapid detection and genotyping of SARS- CoV-2 variants and monitoring of the emerging variants and their dissemination.
Humans ; COVID-19 ; CRISPR-Cas Systems ; Genotype ; Reproducibility of Results ; Reverse Transcriptase Polymerase Chain Reaction ; SARS-CoV-2/genetics* ; RNA ; COVID-19 Testing

Objective:To analyze and reveal the genetic evolution and variation of influenza viruses in cases of co-infection in Guangdong Province.Methods:Throat swab samples were collected from four cases of H1N1pdm and H3N2 co-infection for viral isolation. The isolated strains were subjected to antigen analysis and neuraminidase inhibitor susceptibility test. High-throughput sequencing was used to detect the sequences of strains in three throat swab samples and one virus strain, and then genetic variations were analyzed.Results:Four influenza viruses were isolated with one strain of H1N1pdm and three of H3N2 subtype, and all of them were genetically similar to the vaccine strain in 2022-2023. The HA genes of H1N1pdm and H3N2 strains belonged to clade 6B.1A.5a.2a and 2a.3a.1, respectively. The isolated strains belonged to the same clade as the strains prevalent in Guangdong during the same period. No drug-resistant variations were detected in N1 or N2 gene, and the isolated strains were sensitive to oseltamivir and zanamivir.Conclusions:H1pdm subtype had stronger replication ability than H3 subtype in the influenza viruses isolated from co-infected cases. H1N1pdm and H3N2 subtype influenza viruses were genetically similar to the strains circulating in Guangdong at the same time. The isolated H1N1pdm and H3N2 strains were sensitive to both oseltamivir and zanamivir, indicating that they could continue to be used in the treatment of influenza virus infections caused by one or two genotypes.

Abstract In order to achieve the goal of reducing the smoking rate among Chinese adults at ages of 15 years and above to below 20% by 2030, smoking cessation services require to be reinforced. The current Chinese smoking cessation service system includes short-term smoking cessation interventions and smoking cessation hotlines at the population level, and smoking cessation clinics at the individual level; however, these smoking cessation services suffer from problems of low awareness, low accessibility and low utilization rate. Based on the publications pertaining to smoking cessation services and released policy documents in China until June 2022, this review analyzes the current status, problems and causes of smoking cessation services, and proposes suggestions for improving the smoking cessation service system construction in China.