BACKGROUND:In consideration of the food safety and ecological safety of transgenic plants,the retention of marker genes is the primary safety issue affecting transgenic plants. OBJECTIVE:Based on the principle of immune caries prevention,our research team successfully constructed the plant anti-caries vaccine fusion gene expression vector pCAMBIA-E8-APB-DOCK8 for these two caries causing virulence factors surface protein and glucosyltransferase,which provides a basis for the research and development of transgenic plant vaccine. METHODS:In this study,the selective marker genes Km and GUS in the plant caries vaccine fusion gene expression vector pCAMBIA-E8-APB-DOCK8 were removed by DNA recombination technology through a series of steps such as DNA fragment separation,connection,transformation,clone detection,and sequencing. RESULTS AND CONCLUSION:The efficiency of marker gene removal was 99%.This study has laid a good experimental foundation for the safe production of transgenic plant vaccine against dental caries,and also provided ideas for the construction of other plant vaccine vectors.

BACKGROUND:The application of orthodontic force triggers autophagy in the periodontal tissue via diverse signaling pathways,augmenting or attenuating the activity of relevant cell types such as periodontal ligament cells,osteocytes,osteoclasts,and osteoblasts,thus facilitating the process of periodontal remodeling. OBJECTIVE:To review the research progress in orthodontic force mediated autophagy in periodontal tissue and its impact on orthodontic tooth movement. METHODS:The PubMed,Web of Science,China Biology Medicine disc and CNKI were searched for literature published from 2010 to 2023 to summarize the progress in orthodontics-related autophagy.And 76 papers were finally included in the analysis and discussion. RESULTS AND CONCLUSION:Orthodontic force can trigger a series of biochemical signal changes through periodontal mechanical receptors and aseptic inflammation they cause,leading to autophagy in periodontal tissue.Subsequently,autophagy generates corresponding feedback through cascaded amplified signaling pathways such as Phosphoinositide 3-kinase/protein kinase B,Hippo,and mitogen-activated protein kinase pathways,promoting periodontal tissue remodeling and ultimately achieving tooth movement and stability.Orthodontic force-induced autophagy can differentially regulate bone resorption on the tooth pressure side and bone formation on the tension side.Related targets have good prospects in the clinical application of orthodontic treatment.Orthodontics and autophagy have complex mechanisms.However,existing research has only focused on exploring the role of autophagy in orthodontic tooth movement.Further exploration is needed to investigate the mutual regulatory effects between autophagy and orthodontic tooth movement,as well as the interactions between upstream mechanical receptors and signaling pathways involved in related pathways.

Ankylosing spondylitis (AS) combined with lower cervical fracture is often categorized into unstable fracture, with a high incidence of neurological injury and a high rate of disability and morbidity. As factors such as shoulder occlusion may affect the accuracy of X-ray imaging diagnosis, it is often easily misdiagnosed at the primary diagnosis. Non-operative treatment has complications such as bone nonunion and the possibility of secondary neurological damage, while the timing, access and choice of surgical treatment are still controversial. Currently, there are no clinical practice guidelines for the treatment of AS combined with lower cervical fracture with or without dislocation. To this end, the Spinal Trauma Group of Orthopedics Branch of Chinese Medical Doctor Association organized experts to formulate Clinical guidelines for the treatment of ankylosing spondylitis combined with lower cervical fracture in adults ( version 2024) in accordance with the principles of evidence-based medicine, scientificity and practicality, in which 11 recommendations were put forward in terms of the diagnosis, imaging evaluation, typing and treatment, etc, to provide guidance for the diagnosis and treatment of AS combined with lower cervical fracture.

Low-intensity pulsed ultrasound(LIPUS)was first introduced in clinic for the treatment of fracture because of its safety,ef-fectiveness and non-trauma.Then,a large number of subsequent studies have used LIPUS as a synergistic factor in bone,cartilage,joint,cancer treatment and oral clinical research,jointly revealing the powerful synergistic effect of LIPUS.This paper summarizes the synergistic effect of LIPUS in the above research fields,in order to broaden the possible clinical application scope of LIPUS.

China prospective multicenter birth cohort (Prospective Omics Health Atlas birth cohort, POHA birth cohort) study was officially launched in 2022. This study, in collaboration with 12 participating units, aims to establish a high-quality, multidimensional cohort comprising 20 000 naturally conceived families and assisted reproductive families. The study involves long-term follow-up of parents and offspring, with corresponding biological samples collected at key time points. Through multi-omics testing and analysis, the study aims to conduct multi-omics big data research across the entire maternal and infant life cycle. The goal is to identify new biomarkers for maternal and infant diseases and provide scientific evidence for risk prediction related to maternal diseases and neonatal health.

Abstract: Objective To understand the morphological characteristics and ultrastructure of the dominant species of Rhipicephalus sanguineus in Hainan at different developmental stages, and provide theoretical basis for the identification of the lineage and control of Rhipicephalus sanguineus. Methods The external morphology of different developmental stages of the dominant species of Rhipicephalus sanguineus, including larva, nymph and adult tick in Hainan were observed by scanning electron microscope. Results The division between each segment of larva pedipalps was not obvious, and setae was serrated; dental formula type 2 | 2; 3 pairs of podomere; a pair of setae on the anal valve; none of anal groove, spiracular plate, porous area and genital aperture. There was a clear boundary at the beginning of each segment of nymph pedipalps; dental formula type 2 | 2; 4 pairs of podomere; 3 pairs of setae on the anal valve; anal groove; none of porous area and genital aperture. The male adult tick's trichotheca are covered by the pedipalps, and the whole bristles are conical; dental formula type 3 | 3; 4 pairs of podomere; anal groove and paraprocts; 7 setae on the anal valve; genital aperture was oval. The female of adult tick can be distinguished by dental formula 3 | 3; pairs of podomere; porous areas with 3 short setae; anal groove; 4 pairs of setae and 2 pores on the anal valve; genital pore was broadly U-shaped. In addition, the male adult's scutum occupies almost the entire dorsal surface, the basis capituli of larva, nymph and adult tick all were hexagonal, and the existence of Haller's organ was found on the first pair of legs. Conclusions Scanning electron microscopy observation of the different developmental stages of R.sanguineus revealed clear morphological features, preliminarily suggesting that R.sanguineus in Hainan Province may belong to the tropical lineage, which provide a certain experimental basis for the identification of the tick and the comprehensive prevention and control of local tick-borne diseases.

OBJECTIVE: To explore the role of endoplasmic reticulum ryanodine receptor 1 (RyR1) expression and phosphorylation in sepsis- induced diaphragm dysfunction. METHODS: Thirty SPF male SD rats were randomized equally into 5 groups, including a sham-operated group, 3 sepsis model groups observed at 6, 12, or 24 h following cecal ligation and perforation (CLP; CLP-6h, CLP-12h, and CLP-24h groups, respectively), and a CLP-24h group with a single intraperitoneal injection of KN- 93 immediately after the operation (CLP-24h+KN-93 group). At the indicated time points, diaphragm samples were collected for measurement of compound muscle action potential (CMAP), fatigue index of the isolated diaphragm and fitted frequencycontraction curves. The protein expression levels of CaMK Ⅱ, RyR1 and P-RyR1 in the diaphragm were detected using Western blotting. RESULTS: In the rat models of sepsis, the amplitude of diaphragm CMAP decreased and its duration increased with time following CLP, and the changes were the most obvious at 24 h and significantly attenuated by KN-93 treatment (P < 0.05). The diaphragm fatigue index increased progressively following CLP (P < 0.05) irrespective of KN- 93 treatment (P>0.05). The frequency-contraction curve of the diaphragm muscle decreased progressively following CLP, and was significantly lower in CLP-24 h group than in CLP-24 h+KN-93 group (P < 0.05). Compared with that in the sham-operated group, RyR1 expression level in the diaphragm was significantly lowered at 24 h (P < 0.05) but not at 6 or 12 following CLP, irrespective of KN-93 treatment; The expression level of P-RyR1 increased gradually with time after CLP, and was significantly lowered by KN-93 treatment at 24 h following CLP (P < 0.05). The expression level of CaMKⅡ increased significantly at 24 h following CLP, and was obviously lowered by KN-93 treatment (P < 0.05). CONCLUSION Sepsis causes diaphragmatic dysfunction by enhancing CaMK Ⅱ expression and RyR1 receptor phosphorylation in the endoplasmic reticulum of the diaphragm.
Rats ; Male ; Animals ; Diaphragm/metabolism* ; Ryanodine Receptor Calcium Release Channel/metabolism* ; Rats, Sprague-Dawley ; Phosphorylation ; Muscle Contraction/physiology* ; Endoplasmic Reticulum ; Sepsis/metabolism*

Ankylosing spondylitis (AS) combined with spinal fractures with thoracic and lumbar fracture as the most common type shows characteristics of unstable fracture, high incidence of nerve injury, high mortality and high disability rate. The diagnosis may be missed because it is mostly caused by low-energy injury, when spinal rigidity and osteoporosis have a great impact on the accuracy of imaging examination. At the same time, the treatment choices are controversial, with no relevant specifications. Non-operative treatments can easily lead to bone nonunion, pseudoarthrosis and delayed nerve injury, while surgeries may be failed due to internal fixation failure. At present, there are no evidence-based guidelines for the diagnosis and treatment of AS combined with thoracic and lumbar fracture. In this context, the Spinal Trauma Academic Group of Orthopedics Branch of Chinese Medical Doctor Association organized experts to formulate the Clinical guideline for the diagnosis and treatment of adult ankylosing spondylitis combined with thoracolumbar fracture ( version 2023) by following the principles of evidence-based medicine and systematically review related literatures. Ten recommendations on the diagnosis, imaging evaluation, classification and treatment of AS combined with thoracic and lumbar fracture were put forward, aiming to standardize the clinical diagnosis and treatment of such disorder.

The acute combination fractures of the atlas and axis in adults have a higher rate of neurological injury and early death compared with atlas or axial fractures alone. Currently, the diagnosis and treatment choices of acute combination fractures of the atlas and axis in adults are controversial because of the lack of standards for implementation. Non-operative treatments have a high incidence of bone nonunion and complications, while surgeries may easily lead to the injury of the vertebral artery, spinal cord and nerve root. At present, there are no evidence-based Chinese guidelines for the diagnosis and treatment of acute combination fractures of the atlas and axis in adults. To provide orthopedic surgeons with the most up-to-date and effective information in treating acute combination fractures of the atlas and axis in adults, the Spinal Trauma Group of Orthopedic Branch of Chinese Medical Doctor Association organized experts in the field of spinal trauma to develop the Evidence-based guideline for clinical diagnosis and treatment of acute combination fractures of the atlas and axis in adults ( version 2023) by referring to the "Management of acute combination fractures of the atlas and axis in adults" published by American Association of Neurological Surgeons (AANS)/Congress of Neurological Surgeons (CNS) in 2013 and the relevant Chinese and English literatures. Ten recommendations were made concerning the radiological diagnosis, stability judgment, treatment rules, treatment options and complications based on medical evidence, aiming to provide a reference for the diagnosis and treatment of acute combination fractures of the atlas and axis in adults.

Objective　To investigate the biological characteristics of the serine protease inhibitor 15 (RmS-15) protein of Rhipicephalus microplus, and to perform molecular cloning and prokaryotic expression. Methods RmS-15 gene was amplified and sequenced to construct a phylogenetic tree to understand its evolutionary relationships. Bioinformatic tools were used to analyze the physicochemical properties, signal peptide, secondary and tertiary structure of the RmS-15 protein. In addition, DNA star and ABCpred were used to predict potential B-lymphocyte antigenic epitopes of RmS-15 protein, and potential T-lymphocyte antigenic epitopes were predicted by SYF-PEITHI and IEDB. Finally, the RmS-15 recombinant protein of Rhipicephalus microplus was obtained using the E.coli prokaryotic expression system and purified. Results The RmS-15 gene with 1 212 bp was successfully amplified, encoding a protein comprising 403 amino acids. Phylogenetic tree results indicated high conservation of the RmS-15 protein among different tick amino acid sequences, with the closest phylogenetic relationships to the Rhipicephalus haemaphysaloides and the Rhipicephalus sanguineus. The results of physicochemical property analysis showed a 20-amino acid signal peptide at the N-terminus of the RmS-15 protein, with a relative molecular weight and theoretical isoelectric point of 44 000 and 6.68, respectively. The protein showed an average hydrophilicity of 0.001, classifying it as a stably hydrophilic protein. The results of antigenicity analysis showed that the dominant fragments of B-cell antigenic epitopes of RmS-15 protein were 102-112 aa,147-152 aa and 207-215 aa, and the dominant fragments of T-cell antigenic epitopes were 3-11 aa, 6-14 aa, 27-33 aa, 34-37 aa. Protein expression results showed that RmS-15 protein exhibited high expression levels in the supernatant after induction with 0.8 mmol/L IPTG at 16 ℃ for 24 hours and reached a successful purification with a single band of the protein molecular weight of 44 000. Conclusions The method of prokaryotic expression and purification of RmS-15 protein from Rhipicephalus microplus was successfully established. Bioinformatics analysis demonstrated its strong antigenicity, suggesting its potential to develop as a candidate vaccine for ticks.