Objective : To investigate the causes of false negative results in the detection of Streptococcus agalactiae using fluorescent quantitative PCR(qPCR) targeting the CAMP factor gene(cfb),and to perform a comprehensive analysis of the associated molecular mechanisms. Methods: A total of 76 vaginal secretion samples were evaluated using both qPCR based on cfb gene and bacterial culture methods. Four suspicious strains exhibiting negative qPCR results but positive culture findings were identified using matrix-assisted laser desorption ionization time-of-flight mass spectrometry(MALDI-TOF MS),latex agglutination antigen detection,and the CAMP test. Whole genome analysis was conducted utilizing the MGI DNBSEQ-T7 and Nanopore-PromethION 48 sequencing platforms. Phylogenetic and circular evolutionary trees were constructed using the 16S rRNA gene for strain verification. Multilocus sequence typing(MLST) was conducted,and cfb sequences were aligned and analyzed based on spliced sequences and original data. Specific primers targeting the cfb gene were designed for full-length amplification,followed by verification through agarose gel electrophoresis. Results: The four strains identified as suspicious were classified as S. agalactiae through MALDI-TOF MS,antigen detection,and 16S rRNA gene analysis,with MLST typing indicating ST-862. Phenotypic analysis revealed a negative CAMP test. Whole genome sequence alignment failed to detect the cfb gene or any homologous sequences,and molecular testing confirmed the absence of cfb gene PCR amplification products,thereby confirming its complete deletion. Conclusion This deletion is identified as the molecular mechanism responsible for the false negative qPCR detection of S. agalactiae when targeting this specific gene.It is recommended that the qPCR detection targeting a single cfb gene has limitation,and this may impact clinical diagnosis and treatment decisions. This limitation warrants carefulconsideration.

Objective: To investigate the hemolytic phenotypes of Staphylococcus aureus clinical isolates． Methods: The hemolytic phenotypes of 105 Staphylococcus aureus isolates were analyzed and summarized using the three-point inoculation method．Real-time fluorescence quantitative PCR was used to measure the mRNA expression levels of four hemolysin genes (hla，hlb，hlc，and hld) ; The VITEK 2 GP639 antimicrobial susceptibility card was used to detect resistance to commonly used antibiotics ; DNA gel electrophoresis was performed to determine the prevalence of the mecA，sea，tst，and pvl genes ; The microtiter plate crystal violet staining method was used to assess biofilm formation ability ; The CCK-8 assay was used to evaluate cytotoxicity against macrophages． Results: Seven hemo- lytic phenotypes were identified among the Staphylococcus aureus clinical isolates． Differences were found among Staphylococcus aureus clinical isolates with different hemolytic phenotypes in terms of mRNA expression levels of he- molysin genes，antibiotic resistance，virulence gene prevalence，biofilm formation ability，and cytotoxicity to mouse macrophages (P ＜0. 05 ) ． Conclusion Staphylococcus aureus clinical isolates exhibit diverse hemolytic pheno- types，which should be a focus across multiple dimensions，including microbiological testing，clinical treatment， and nosocomial infection prevention and control．

In recent years,viral vector-mediated chimeric antigen receptor(CAR)cell therapy has emerged as a prominent topic in the field of cancer treatment and has demonstrated significant clinical efficacy in patients with hematological malignancies.However,the permanent expression of CAR induced by this therapy may give rise to severe adverse reactions.In contrast,messenger RNA(mRNA)chimeric antigen receptor therapy is anticipated to become a new generation of safer and more effective treatments due to its lack of insertional mutagenesis and minimal risk of off-tar-get toxicity.This therapeutic approach involves transfecting effector immune cells with CAR-en-coding mRNA to elicit an immune response within the body.It has been employed for treating B-cell malignancies,lymphoma,acute myelogenous leukemia(AML),and other cancers.The continuous optimization of mRNA purification,modification,and transfection technology enhances CAR ex-pression durability,while increasing killing efficiency and expanding the range of action.This pa-per reviews the research advances on CAR-encoding mRNA delivery,its application and advan-tages in cancer therapies.

Nonobstructive azoospermia (NOA), one of the most severe types of male infertility, etiology often remains unclear in most cases. Therefore, this study aimed to detect four biallelic detrimental variants (0.5%) in the minichromosome maintenance domain containing 2 ( MCMDC2 ) genes in 768 NOA patients by whole-exome sequencing (WES). Hematoxylin and eosin (H&E) demonstrated that MCMDC2 deleterious variants caused meiotic arrest in three patients (c.1360G>T, c.1956G>T, and c.685C>T) and hypospermatogenesis in one patient (c.94G>T), as further confirmed through immunofluorescence (IF) staining. The single-cell RNA sequencing data indicated that MCMDC2 was substantially expressed during spermatogenesis. The variants were confirmed as deleterious and responsible for patient infertility through bioinformatics and in vitro experimental analyses. The results revealed four MCMDC2 variants related to NOA, which contributes to the current perception of the function of MCMDC2 in male fertility and presents new perspectives on the genetic etiology of NOA.
Humans ; Male ; Azoospermia/genetics* ; Meiosis/genetics* ; Spermatogenesis/genetics* ; Adult ; Exome Sequencing ; Microtubule-Associated Proteins/genetics* ; Alleles ; Infertility, Male/genetics*

Cervical spinal cord injury without fracture-dislocation (CSCIWFD) is referred to as a special type of cervical spinal cord injury characterized by traumatic spinal cord dysfunction and no significant bony structural abnormalities on imagines. Duo to the high risk of missed diagnosis during the initial consultation, CSCIWFD may lead to progressive neurological deterioration or even complete paralysis, severely impacting patients′ prognosis. Currently, there are no established consensuses over the diagnosis and treatment of CSCIWFD, such as the lack of evidence-based standards for indications of non-surgical treatment and risk of secondary neurological injury, as well as debates over the optimal timing for surgical intervention and indications for different surgical approaches. To address these issues, the Spine Trauma Group of the Orthopedic Branch of the Chinese Medical Doctor Association organized experts in the relevant fields to formulate Diagnosis and treatment guideline for acute cervical spinal cord injury without fracture- dislocation in adults ( version 2025) . Based on evidence-based medicine and the principles of scientific rigor and clinical applicability, the guidelines proposed 11 recommendations covering terminology, diagnosis, evaluation treatment, and rehabilitation, etc., aiming to standardize the management of CSCIWFD.

With the rapid development of competitive sports, the incidence of anterior cruciate ligament (ACL) injury is on the rise. Such injuries may shorten athletes′ career and lead to other long-term adverse consequences. Although athletes generally recover well after ACL reconstruction, many still struggle to return to their pre-injury performance levels. Advances in the understanding of ACL anatomy and injury mechanisms, along with the evolution of surgical techniques and rehabilitation methods, have provided more individualized and tailored options for athletes following ACL injuries. However, there is currently no consensus in China regarding surgical and rehabilitation strategies for competitive athletes aiming to return to sports after ACL injuries. To this end, the Sports Medicine Committee of the Chinese Research Hospital Association and the Editorial Board of the Chinese Journal of Trauma jointly formulated the Expert consensus on surgical treatment and rehabilitation for competitive sports athletes returning to sports after anterior cruciate ligament injury ( version 2025), and presented 14 recommendations covering surgical indications, preoperative rehabilitation, surgical timing, surgical strategies and postoperative rehabilitation strategies, aiming to improve the surgical treatment and rehabilitation system for ACL injuries in competitive athletes and facilitate their return to high-level sports performance after injury.

Vertebral refracture following percutaneous vertebral augmentation (PVA) is commonly seen in elderly patients with osteoporotic thoracolumbar compression fractures (OTLCF). It can lead to recurrent pain, loss of vertebral height, progression of kyphosis, and even neurological dysfunction, significantly impairing patients′ quality of life. Current diagnosis and treatment face multiple challenges, including high misdiagnosis rate, difficulty in choosing between surgical and non-surgical treatment options, lack of standardized surgical protocols, interference from intralesional bone cement during procedures, inadequate stability of internal fixation in osteoporotic bone, and suboptimal compliance of anti-osteoporotic therapy. Establishing a standardized diagnostic and therapeutic framework is urgently needed. To standardize the management process and improve outcomes for vertebral refractures after PVA in elderly OTLCF patients, Spinal Trauma Group of the Orthopedic Branch of Chinese Medical Doctor Association organized experts in the field to develop Guideline for the diagnosis and treatment of vertebral refracture after percutaneous vertebral augmentation in elderly patients with osteoporotic thoracolumbar compression fractures ( version 2025), based on current literature and clinical experience, and adhering to principles of scientific rigor and clinical applicability. A total of 11 recommendations were proposed, encompassing diagnosis, treatment, and rehabilitation of vertebral refracture after PVA in elderly patients with OTLCF, aiming to provide a foundation for a standardized management.

For middle-aged and elderly patients with conditions such as spinal fractures and degenerative spinal diseases, spinal internal fixation is a core surgical procedure for reconstructing spinal stability, heavily relying on the biomechanical stability provided by pedicle screw systems. Whereas, these patients are often complicated by osteoporosis that can significantly compromise the stability of the bone-pedicle screw interface, leading to a marked increase in pedicle screw loosening and surgical failure rates. The bone cement-augmented pedicle screw technique, which involves injecting bone cement into the vertebral body or screw trajectory to optimize the mechanical properties of the bone-pedicle screw composite, has been proven to significantly enhance fixation strength and effectively prevent screw-related failures, thereby reducing the incidence of internal fixation failure in high-risk populations undergoing spinal fusion. However, the widespread clinical application of this technique has faced challenges such as inaccurate clinical decision-making (indication and contraindication selection), non-standardized operative practices, and insufficient awareness of complication prevention, resulting in considerable variability in clinical outcomes and even severe complications. To address this, Prof. Luo Fei from First Affiliated Hospital of Army Medical University initiated the project and the Chinese Association Orthopaedic Surgeons organized relevant experts to develop the Evidence-based clinical practice guideline for bone cement-augmented pedicle screw technique ( version 2025), based on current evidence. The guidelines put forward 8 recommendations regarding the clinical value, scope of application, and operational standards of the technique, aiming to provide evidence-based medical support and technical standardization for clinical decision-making.

Objective:To investigate the causes and management strategies of anesthetic complications during percutaneous spinal endoscopic surgery under local anesthesia.Methods:A total of 16 800 patients (8 625 males and 8 175 females) who underwent percutaneous spinal endoscopic surgery under local anesthesia (including intravenous basic anesthesia) in Tianjin Hospital, Shandong Public Health Clinical Center and Hebei General Hospital from February 2012 to February 2023 were retrospectively analyzed. The average age was 45.3±21.6 years (range, 12-84 years). There were 220 cases of posterior cervical keyhole endoscopic surgery, 50 cases of thoracic transforaminal endoscopic surgery, 70 cases of thoracic posterior interlaminar endoscopic surgery, 11 670 cases of lumbar transforaminal endoscopic surgery, and 4 790 cases of lumbar posterior interlaminar endoscopic surgery. The occurrence time, clinical manifestations, management of intraoperative anesthesia complications were recorded, as well as surgical segments, puncture sites, complication symptoms, signs, outcome and prognosis.Results:All patients received percutaneous water-mediated uniaxial spinal endoscopic surgery under local anesthesia. There were 9 patients experienced anesthesia complications, including 6 cases of epidural diffusion of anesthetics and 3 cases of anesthetics mistakenly entering the subarachnoid space. There were 4 males and 5 females, aged 48.4±18.2 years (range, 28-84 years). There were 1 case of T 12L 1 disc herniation, 1 case of C 5-6 disc herniation, 3 cases of L 4-5 disc herniation and 4 cases of L 5S 1 disc herniation. Surgical segments and procedures: 1 case of C 5-6 posterior Keyhole endoscopic surgery, 1 case of T 12L 1 transforaminal endoscopic surgery, 2 cases of L 4-5 transforaminal endoscopic surgery, 1 case of L 4-5 interlaminar endoscopic surgery, and 4 cases of L 5S 1 interlaminar endoscopic surgery. Anesthesia complications all appeared 5-10 min after injection of local anesthetics, with symptoms of decreased oxygen saturation, decreased blood pressure, altered consciousness, and sensory and motor dysfunction of limbs. 6 patients with epidural diffusion of anesthetics recovered completely after symptomatic treatment in 5 cases, and 1 case was left with foot drop. Three patients with anesthetics mistakenly entering the subarachnoid space were immediately converted to the supine position, of which one recovered by mask oxygenation; 1 patient improved after emergency tracheal intubation, rehydration, and application of vasoconstrictive medications; and 1 patient developed multiple complications such as multiorgan failure, rhabdomyolysis, and sepsis after tracheal intubation, and recovered at 3 months after surgery with symptomatic treatment. Conclusions:Epidural diffusion and entering into subarachnoid space of anesthetics are serious complications of local anesthesia in percutaneous spinal endoscopic surgery. In addition to sensory and motor dysfunction of the limbs, the functions of the respiratory and circulatory systems can also be affected. It is necessary to be alert to the occurrence of anesthesia-related complications during operation and early identification and treatment.

Body weight abnormalities, including overweight, obesity, and underweight, have become a dual public health challenge in Chinese adults: overweight and obesity lead to a variety of chronic complications, while underweight increases the risks of malnutrition, sarcopenia, and organ dysfunction. To systematically address these issues, multidisciplinary experts in endocrinology, sports science, nutrition, and psychiatry from various regions have held multiple weight management seminars. Based on the latest epidemiological data and clinical evidence, they expanded the guideline to include assessment and intervention strategies for underweight, in addition to the core content of obesity management. This guideline outlines the etiological mechanisms, evaluation methods, and multidimensional management strategies for overweight and obesity, covering key areas such as diagnosis and assessment, medical nutrition therapy, exercise prescription, pharmacological intervention, and psychological support. It is intended to provide a scientific and standardized approach to weight management across the adult population, aiming to curb the rising prevalence of obesity, mitigate complications associated with abnormal body weight, and improve nutritional status and overall quality of life.