Since its emergence in the 1980s, the human immunodeficiency virus (HIV) has caused a global pandemic, posing a severe threat to human life and health as well as social development. Although pre-exposure prophylaxis (PrEP) effectively curbs HIV transmission and antiretroviral therapy (ART) significantly extends the lifespan of patients, vaccines remain a pivotal tool for blocking transmission and ending the pandemic. The high genetic variability of HIV-1, the glycan shield of its envelope glycoproteins, and the long-term persistence of latent reservoirs have repeatedly led to bottlenecks in traditional vaccine strategies. In recent years, mRNA technology has offered a novel approach to addressing these challenges, leveraging advantages such as sequence programmability, short production cycles, native conformational expression of antigens, and self-adjuvant effects. In recent years, mRNA vaccine technology has emerged as a transformative solution to longstanding vaccinology challenges, characterized by its sequence programmability, rapid production cycles, native conformational antigen expression, and intrinsic self-adjuvanting properties. Unlike traditional platforms reliant on pathogen culture or recombinant proteins, mRNA vaccines can be expeditiously designed and updated based solely on viral genomic sequences. Lipid nanoparticle (LNP)-encapsulated mRNA facilitates endogenous antigen expression and presentation, simultaneously eliciting potent humoral and cellular immune responses. Within this landscape, self-amplifying mRNA (saRNA) further extends in vivo antigen expression to enhance the persistence of immune responses. Moreover, the LNP delivery system not only protects mRNA from degradation and mediates endosomal escape but also synergizes with mRNA to optimize immune activation via self-adjuvant effects. Importantly, mRNA platforms circumvent the pre-existing immunity associated with viral vectors and the genomic integration risks of DNA vaccines, positioning them as a cornerstone for global pandemic preparedness. This review systematically delineates recent advances in mRNA technology for HIV-1 vaccine development, focusing on four pivotal research frontiers. First, mRNA innovations building upon the RV144 trial optimize antigens through codon modification and multivalent designs to induce more durable and broad-spectrum immunity. Second, particulate mRNA vaccine strategies, utilizing virus-like particles (VLPs) and ferritin nanoparticles, achieve in situ antigen self-assembly, significantly enhancing B cell activation and reducing infection risks in non-human primate models. Third, germline-targeting mRNA vaccines address the low-affinity barrier of broadly neutralizing antibody (bNAp) precursors, efficiently activating rare precursor B cells and promoting affinity maturation. Fourth, therapeutic mRNA vaccines offer unique advantages for an HIV functional cure; combining immunogens with mRNA-encoded adjuvants potentiates cellular immunity, while LNP-mediated “shock-and-kill” strategies specifically activate latent reservoirs to guide immune clearance. Comparative analyses with traditional platforms reveal that mRNA technology redefines antigen production and presentation, simulating chronic infection through sustained expression and enabling dual-pathway presentation via endogenous synthesis. Furthermore, we explore the mechanistic innovations of mRNA vaccines in inducing bNAps: sustained in vivo production prolongs the activation window for precursor B cells and maintains germinal center (GC) reactions; endogenously expressed antigens adopt native conformations to expose conserved epitopes; and self-adjuvanting effects modulate the functions of antigen-presenting cells (APCs) and follicular helper T cells (Tfh), driving somatic hypermutation and affinity maturation. We also address critical clinical translation challenges, including immune durability, adaptability to special populations, and large-scale LNP manufacturing, while proposing targeted optimization strategies. In conclusion, this review establishes a theoretical framework for utilizing mRNA technology to overcome HIV-1 immune escape, transitioning from a descriptive paradigm to a problem-solving-based synthesis of evidence. By integrating preclinical and early clinical data, we bridge the gap between basic design and translational verification. mRNA technology is poised to become a central pillar inHIV-1 prevention and therapy, providing a robust toolset to achieve the global goal of ending the AIDS pandemic and offering a blueprint for vaccine development against other recalcitrant infectious diseases.

Objective:To investigate spinal cord morphological changes and risk factors in upper cervical spine surgery using C 2 medial "in-out-in" pedicle screws in patients with atlantoaxial dislocation (AAD) and high-riding vertebral artery (HRVA). Methods:A retrospective analysis was conducted on 41 patients with AAD who underwent C 2 medial "in-out-in" pedicle screw implantation at our hospital between January 2019 and December 2023. The cohort included 12 males and 29 females, with a mean age of 47.6±10.3 years (range: 18-68 years). Among them, 30 patients received unilateral C 2 medial "in-out-in" screws, while 11 patients received bilateral screws. All patients underwent posterior reduction and internal fixation. Measurements included C 2 pedicle height, C 2 pedicle width, C 2 horizontal spinal canal width, screw invasion distance into the spinal canal, and spinal canal invasion rate based on CT findings. MRI evaluations included spinal canal-dura mater distance, dura mater-spinal cord distance, spinal canal-spinal cord distance, and spinal cord cross-sectional area. The change rate of spinal cord cross-sectional area was calculated and categorized into >20% decrease group (decreasing group) and ≤20% decrease group (non-decreasing group). Postoperative outcomes were assessed using the Visual Analogue Scale (VAS) and Japanese Orthopedic Association (JOA) scores. Results:The mean operative time was 165.5±30.1 minutes (range: 120-220 minutes). No spinal cord vascular injuries or severe complications were observed. However, five patients experienced cerebrospinal fluid (CSF) leakage, requiring delayed removal of drainage tubes, but their wounds healed successfully. Follow-ups (range: 6-24 months) showed no loosening of internal fixation, fractures, or bone graft nonunion, with a 100% bone fusion rate at 6 months postoperatively. VAS scores improved significantly from a preoperative median of 5.0 (3.5, 6.0) (range: 1-7) to 2.0 (2.0, 3.0) at 3 months and 1.0 (1.0, 1.0) at 6 months ( Z=36.716, P< 0.001). JOA scores improved from 10.0 (9.0, 12.0) (range: 4-14) preoperatively to 13.0 (11.0, 13.5) at 3 months and 14.0 (12.0, 15.0) at 6 months ( Z=67.093, P<0.001). The height of C 2 pedicle was 5.50±1.78 mm, the width of C 2 pedicle was 2.27±1.23 mm, the width of C 2 horizontal spinal canal was 23.76±4.91 mm, the spinal canal-dura mater distance was 3.08±0.85 mm, dura mater-spinal cord distance was 3.23±0.85 mm, the spinal canal-spinal cord distance was 6.31±1.11 mm, the distance of screw invasion into the spinal canal was 2.80±1.54 mm, the rate of spinal canal invasion was 12.1%±6.8%, the preoperative spinal cord cross-sectional area was 69.81±13.27 mm 2, and the postoperative spinal cord cross-sectional area was 68.81±13.94 mm 2. Based on spinal cord cross-sectional area changes, 32 patients were classified into the non-decreasing group, and 9 patients into the decreasing group. The intraclass correlation coefficient (ICC) values ranged from 0.733 to 0.984, indicating high measurement reliability. There were significant differences in the proportion of C 2 bilateral medial "in-out-in" screws (χ 2=6.903, P=0.009), the incidence of CSF leakage (χ 2=15.391, P<0.001), the distance of screw invasion into the spinal canal ( t=4.990, P<0.001) and the rate of spinal canal invasion ( t=4.045, P<0.001) in the decreasing group versus the non-decreasing group. The JOA scores of the non-decreasing group were significantly higher at 3 and 6 months postoperatively compared to the decreasing group ( P<0.05). No other parameters showed significant differences between the groups. Binary logistic regression analysis identified spinal canal invasion rate [ OR=1.963, 95% CI (1.010, 3.817), P=0.047] as an independent risk factor for spinal cord cross-sectional area reduction. The Jordan index was 0.875, with a spinal canal invasion rate threshold of 14.18%, a sensitivity of 1.000, a specificity of 0.875, and an AUC of 0.983, indicating strong predictive value. Conclusion:The C 2 medial "in-out-in" screw technique provides effective posterior fixation and fusion for AAD patients with HRVA. However, to minimize spinal cord morphological changes and associated risks, the spinal canal invasion rate should be kept below 14.18% when using this technique.

Prolactin, a peptide hormone primarily synthesized and secreted by lactotrophs in the anterior pituitary gland, plays key roles in lactation, reproduction, and immune regulation. Recent research has highlighted its critical involvement in maintaining metabolic homeostasis, with both elevated and deficient levels disrupting metabolic functions such as glucose and lipid metabolism, as well as energy regulation. This study reviews recent advancements in prolactin and its receptors′ role in metabolic regulation, emphasizing its pivotal contribution to metabolic homeostasis.

Objective:To evaluate the diagnostic performance of the 0-2 h high-sensitivity cardiac troponin T (hs-cTnT) cutoff recommended by the guidelines for the rule-out and rule-in diagnosis of suspected non-ST-segment elevation myocardial infarction (NSTEMI) patients of different age groups.Methods:This is a retrospective cohort study. Clinical data of 4 050 suspected NSTEMI patients who visited the Chest Pain Center of Zhongshan Hospital affiliated with Fudan University from January 2020 to December 2021 were retrospectively analyzed. Patients who visited from January 2020 to April 2021 (2 650 patients) were included as derivation cohort, and those who visited from May to December 2021 (1 400 patients) were included as validation cohort. The diagnostic performance of the guideline-recommended hs-cTnT 0-2 h cutoff for the rule-out and rule-in of NSTEMI diagnosis was compared among subgroups of patients aged ≤60, >60-70, and >70 years in the derivation group. Rule-out sensitivity, negative predictive value, and rule-out proportion, rule-in specificity, positive predictive value, and rule-in proportion were assessed. Cutoffs were established for subgroups with relatively lower diagnostic performance and validated in the validation group. Major adverse cardiovascular events (MACE) within 30 days after patient visit were used as the outcome, and survival curves were plotted using Kaplan-Meier curves, log-rank tests were used to analyze the incidence of MACE.Results:The sensitivity for ruled-out NSTEMI using the guideline-recommended 0-2 h cutoff in the subgroups of patients aged ≤60, >60-70, and >70 years in the derivation group was 100%; the negative predictive value was 100%; the ruled-out rates were 47.6% (331/696), 45.9% (491/1 070), and 28.5% (252/884), respectively. The specificity for ruled-in NSTEMI was 88.3%, 90.9%, and 86.4%, respectively; the positive predictive values were 55.3%, 59.3%, and 58.2%, respectively; the ruled-in rates were 22.6% (157/696), 19.5% (209/1 070), and 27.0% (239/884), respectively. With a requirement of sensitivity and negative predictive value >99%, the ruled-out cutoff for the subgroup of patients aged >70 years in the derivation group was established as 0 h hs-cTnT <6 ng/L or 0 h hs-cTnT<22 ng/L and 0-2 h Δhs-cTnT <5 ng/L, which increased the ruled-out rate of the subgroup aged >70 years to 45.6% (403/884). In the validation group, 42.2% (196/465) patients could be ruled-out. The incidence of MACE within 30 days for ruled-out patients aged >70 years using the established cutoff was 0.Conclusion:The diagnostic performance for the ruled-out and ruled-in diagnosis using the guideline-recommended 0-2 h hs-cTnT cutoff are relatively consistent across different age groups, but the ruled-out rate for patients aged >70 years is lower than for those aged ≤60 and >60-70 years. The ruled-out cutoff established in this study can be used to improve diagnostic performance of thus indicator on suspected NSTEMI patients.

Objective:To compare the clinical outcomes of arthroscopic Y-shaped suture suspension fixation and those of open reduction and internal fixation in the treatment of tibial insertion avulsion fractures of the posterior cruciate ligament (PCL).Methods:A retrospective study was conducted to analyze the clinical data from the 23 patients with PCL tibial avulsion fracture who had been admitted to Department of Sports Medicine, The Affiliated Changsha Central Hospital, University of South China, from May 2018 to May 2024. There were 15 males and 8 females with an age of (39.0±15.6) years. By the Meyers-McKeever classification, there were 20 cases of type Ⅱ and 3 cases of type Ⅲ. All patients were divided into 2 groups based on their different surgical schemes: an arthroscopic group of 11 cases treated with Y-shaped suture suspension fixation under arthroscopy and an open group of 12 cases treated with open reduction and internal fixation. The surgical time, intraoperative blood loss, and visual analog scale (VAS) pain scores, Lysholm knee function scores, and International Knee Documentation Committee (IKDC) subjective scores at 1 month after surgery and the last follow-up were recorded and compared between the 2 groups.Results:There was no statistically significant difference in the preoperative general information between the 2 groups, indicating comparability ( P>0.05). The 23 patients were followed up for (13.7±3.8) months after surgery. None of the patients experienced complications like incision infection, neurovascular damage, nonunion or delayed union of fracture. In the arthroscopic group, the surgical time was (97.1±11.1) min, significantly shorter than that in the open group [(64.7±9.0) min], and the intraoperative blood loss (14.9±6.6) mL, significantly less than that in the open group [(29.4±13.2) mL] ( P<0.05). At 1 month after surgery, the VAS pain score, Lysholm knee function score, and IKDC subjective score in the arthroscopic group were, respectively, 3.0 (2.0, 3.0) points, (72.0±4.8) points, and (67.3±2.9) points, all significantly better than those in the open group [5.0 (4.0, 5.0) points, (66.9±4.0) points, and (63.1±2.4) points] ( P<0.05). There was no statistically significant difference in the VAS pain score, Lysholm knee function score, or IKDC subjective score at the last follow-up between the 2 groups ( P>0.05). Conclusions:Both arthroscopic Y-shaped suture suspension fixation and open reduction and internal fixation can achieve satisfactory treatment outcomes for PCL tibial avulsion fractures. The latter shows an advantage in shorter surgical time, but the former advantages in less bleeding volume and better early efficacy.

Objective To summarize the changing prevalence of carbapenem resistance in Enterobacterales based on the data of CHINET Antimicrobial Resistance Surveillance Program from 2015 to 2021 for improving antimicrobial treatment in clinical practice.Methods Antimicrobial susceptibility testing was performed using a commercial automated susceptibility testing system according to the unified CHINET protocol.The results were interpreted according to the breakpoints of the Clinical & Laboratory Standards Institute(CLSI)M100 31st ed in 2021.Results Over the seven-year period(2015-2021),the overall prevalence of carbapenem-resistant Enterobacterales(CRE)was 9.43％(62 342/661 235).The prevalence of CRE strains in Klebsiella pneumoniae,Citrobacter freundii,and Enterobacter cloacae was 22.38％,9.73％,and 8.47％,respectively.The prevalence of CRE strains in Escherichia coli was 1.99％.A few CRE strains were also identified in Salmonella and Shigella.The CRE strains were mainly isolated from respiratory specimens(44.23±2.80)％,followed by blood(20.88±3.40)％and urine(18.40±3.45)％.Intensive care units(ICUs)were the major source of the CRE strains(27.43±5.20)％.CRE strains were resistant to all the β-lactam antibiotics tested and most non-β-lactam antimicrobial agents.The CRE strains were relatively susceptible to tigecycline and polymyxins with low resistance rates.Conclusions The prevalence of CRE strains was increasing from 2015 to 2021.CRE strains were highly resistant to most of the antibacterial drugs used in clinical practice.Clinicians should prescribe antimicrobial agents rationally.Hospitals should strengthen antibiotic stewardship in key clinical settings such as ICUs,and take effective infection control measures to curb CRE outbreak and epidemic in hospitals.

Objective To characterize the changing species distribution and antibiotic resistance profiles of respiratory isolates in hospitals participating in the CHINET Antimicrobial Resistance Surveillance Program from 2015 to 2021.Methods Commercial automated antimicrobial susceptibility testing systems and disk diffusion method were used to test the susceptibility of respiratory bacterial isolates to antimicrobial agents following the standardized technical protocol established by the CHINET program.Results A total of 589 746 respiratory isolates were collected from 2015 to 2021.Overall,82.6％of the isolates were Gram-negative bacteria and 17.4％were Gram-positive bacteria.The bacterial isolates from outpatients and inpatients accounted for(6.0±0.9)％and(94.0±0.1)％,respectively.The top microorganisms were Klebsiella spp.,Acinetobacter spp.,Pseudomonas aeruginosa,Staphylococcus aureus,Haemophilus spp.,Stenotrophomonas maltophilia,Escherichia coli,and Streptococcus pneumoniae.Each microorganism was isolated from significantly more males than from females(P＜0.05).The overall prevalence of methicillin-resistant S.aureus(MRSA)was 39.9％.The prevalence of penicillin-resistant S.pneumoniae was 1.4％.The prevalence of extended-spectrum β-lactamase(ESBL)-producing E.coli and K.pneumoniae was 67.8％and 41.3％,respectively.The overall prevalence of carbapenem-resistant E.coli,K.pneumoniae,Enterobacter cloacae,Pseudomonas aeruginosa,and Acinetobacter baumannii was 3.7％,20.8％,9.4％,29.8％,and 73.3％,respectively.The prevalence of β-lactamase was 96.1％in Moraxella catarrhalis and 60.0％in Haemophilus influenzae.The H.influenzae isolates from children(＜18 years)showed significantly higher resistance rates to β-lactam antibiotics than the isolates from adults(P＜0.05).Conclusions Gram-negative bacteria are still predominant in respiratory isolates associated with serious antibiotic resistance.Antimicrobial resistance surveillance should be strengthened in clinical practice to support accurate etiological diagnosis and appropriate antimicrobial therapy based on antimicrobial susceptibility testing results.

Soy is a vital source of plant carbohydrates.However,it poses significant allergenic risks,particularly to young children and animals.Among the various proteins in soy,β-conglycinin,which con-stitutes approximately 30％of total soy carbohydrates,is a primary allergen.Undigested β-conglycinin can lead to intestinal damage by inhibiting cell growth,disrupting the cytoskeleton,and inducing apopto-sis.It can also enter the lymphatic and circulatory systems,triggering allergic reactions.Conventional ELISA methods for detecting β-conglycinin rely on polyclonal or monoclonal antibodies,which are limited by their large molecular weight,difficulty in accessing the protein core,and sensitivity to acidic and bas-ic conditions.To address these limitations,this study aimed to develop nanobodies(Nbs)against β-con-glycinin.Nbs,derived from the variable regions of heavy-chain antibodies found in camelids,have a mo-lecular weight approximately one-tenth that of conventional antibodies.They offer advantages such as small size,stable structure,high specificity,and strong affinity.A female alpacas was immunized five times using β-conglycinin,which showed a heavy chain antibody potency of 1∶16 000 by ELISA.Pe-ripheral blood lymphocytes were subsequently isolated and total RNA was extracted.The variable region of the heavy-chain antibody was amplified via PCR,and recombinant plasmids were constructed and transformed into the E.coli competency strain ER2738.The resulting library contained about 3.5×108 CFU/mL,which increased to 1.15×1012 PFU/mL after phage rescue,with a 100％Nbs gene insertion rate,indicating high diversity.Its Nbs phage output was significantly enriched by four rounds of solid-phase elution with an enrichment rate of 155.9.Four rounds of solid-phase panning yielded 35 positive clones,all of which shared the same amino acid sequence upon sequencing.The selected Nb was ex-pressed in a prokaryotic system,and its binding ability to β-conglycinin was confirmed using Western blotting and ELISA.The results demonstrated excellent specificity and affinity.This research lays the groundwork for developing a rapid and efficient detection method for β-conglycinin using Nbs,potentially enhancing food safety and allergen management.

To establish a method for detecting Toxoplasma gondii based on recombinant polymerase amplification(RPA)technology and apply it to clinical sample validation of pet cats.Using the 529 repeat sequence of the Toxoplasma gondii gene as the target gene sequence,primers and probes were designed,and the Rep-529 recombinant plasmid was constructed as the standard.A fluorescent RPA reaction system was established.Dilute the plasmid standard 10 times to different concentrations as the detection template for sensitivity testing;Specific testing was conducted using genomic DNA from several parasitic spe-cies,including Toxoplasma gondii,Cryptosporidium,Neosporidium,Trichinella spiralis,Giardia flagellata,Babesia bo-vis and Theileria annulata as templates;Simultaneously,fluorescence RPA and RT-PCR were used to detect 52 positive and 40 negative cats clinical samples,and the coincidence rate of the detection results of the two methods were compared and ana-lyzed.The RPA reaction system was successfully established using PTRep recombinant plasmid as the standard,ToxD-F/ToxD-R as the primer,and RepD-P as the fluorescent probe.The reaction temperature was constant at 39 ℃,the reaction time was 30 minutes,and the detection sensitivity was 1 copy/μL.There is no significant cross reaction with parasites such as Cryptosporidium,Neosporidium,Trichinella spiralis,Giardia,Babesia bovis and Theileria annulata,and the specificity is good.A total of 92 clinical fecal samples from cats were tested,and the positive coincidence rate of fluorescence RPA detection method was higher than that of conventional RT-PCR method(98.08％vs.82.69％),and the difference of the positive rate was not statistically significant(X2=1.392,P＞0.05).The fluorescence RPA detection method for Toxoplasma gondii suc-cessfully established in this study has the characteristics of being fast,sensitive,specific,accurate,and reliable.It can be used as a rapid clinical detection kit for Toxoplasma gondii in cats and other animals,providing new technical support for the subsequent epidemiological monitoring and precise clinical diagnosis of toxoplasmosis in cats,other animals,and humans in the future.

Objective In response to the prevalent issue of insufficient professional quality among nursing staff working in care homes,this study aims to conduct a foundational assessment using the"Medical and Elderly Care Integration Institution Elderly Care Staff Professional Quality Assessment Scale"based on the job competency theory,and then investigate the effective-ness of the"333"tiered training model following the initial assessment.Methods A convenience sampling method was used to select 160 caregivers from care homes in Wuxi.The caregivers were equally divided into an experimental group and a control group.A baseline assessment of professional competence was conducted using the professional quality assessment indicator system before training.The experimental group was treated with a tiered training model and training program,which were proposed using literature research and questionnaire survey methods and according to the results of baseline professional quality assessment.The control group did not undergo tiered training,but took the same training program as in the experimental group.SPSS 21.0 soft-ware was used for analysing the training outcome data.Results By comparison,the experimental group showed significantly re-duced learning time and had significantly higher assessment scores.The satisfaction degree of the elderly people and the caregiv-ers of the experimental group was significantly higher than that of the control group.The satisfaction ratings with training course design,training methods,and information-based teaching in the experimental group were significantly higher than those in the control group.Conclusion Assessing the professional quality of medical and elderly caregivers is essential for tired training.Tiered training is an effective way to improve the quality and efficiency of training,and a complete training organization system and a sound management mechanism are essential guarantees for implementing tiered training.