This study is designed to address the development, synthesis, and screening of non-animal-derived nanoantibody libraries. Furthermore, it seeks to elucidate the impact of framework region selection and complementarity-determining region (CDR) design on the characteristics of synthesized nanoantibody libraries. These investigations aim to establish a robust theoretical and technical foundation for enhancing the efficacy, diversity, and practical applicability of synthetic nanoantibody libraries. In this study, a new framework (IGHV3S65*01-IGHJ4*01) was identified based on the high-throughput sequencing results of natural nanobodies, and degenerate primers were designed based on the frequency of amino acids at each position in the complementarity-determining region (CDR) region to synthesize the coding fragments of nanobodies by overlap PCR. After 40 times of electro-transformation, a single-frame synthesized nanobody library (SS-Library) containing 6×10⁹ clones was obtained, and the titer of the library was demonstrated to be 10¹³ PFU/mL after rescue by the helper phage M13K07. Random 48 single colonies were picked for PCR, which revealed an insertion rate of 95.8%. Sanger sequencing results showed that 38 clones had complete sequences, none of which showed cysteines or stop codons, and no identical sequences appeared, suggesting that the library had higher diversity. The library was screened and validated with three antigens, including bovine serum albumin (BSA), acetylcholinesterase (AchE), and immunoglobulin G (IgG). Finally, 2 nanobodies against BSA, 10 against AchE, and 15 against IgG were obtained. One positive clone of each antigen was singled out for recombinant expression, and the results showed that all the three nanobodies were expressed in a soluble form. The binding activity of recombinantly expressed nanobodies was evaluated using indirect enzyme-linked immunosorbent assay (ELISA) and bio-layer interferometry (BLI). The results demonstrated that the anti-AChE and anti-IgG nanobodies exhibited specific binding to their respective antigens, with affinity constants (KD) of 294 nmol/L and 250 nmol/L, respectively. The nanobody synthetic library preparation method proposed in this study is simple and easy to use with low preference, and it is expected to be a universal nanobody discovery platform for the preparation and development of lead specific nanobodies.
Single-Domain Antibodies/biosynthesis* ; Peptide Library ; Complementarity Determining Regions/immunology* ; Animals

Atypical lipomatous tumor(ALT)is a rare soft tissue sarcoma originating from adipocytic tissue.The clinical manifestations,imaging findings,and pathological examinations of one patient with ALT in the lower extremity were reported to provide reference for the clinical diagnosis and treatment of this disease.The patient was a 64-year-old male who was admitted due to a mass in the left thigh with swelling and pain for 5 years.The physical examination results showed a subcutaneous mass was palpable on the medial side of the left thigh,soft in texture,demonstrating good mobility and positive tenderness.The magnetic resonance imaging(MRI)non-contrast scan showed a well-defined lesion with an intact capsule.The lesion presented high signal intensity on T1-weighted image(T1WI),similar to subcutaneous fat signal;on T2-weighted image(T2WI)-Ideal in-phase images,the lesion showed high signal,while water images showed low signal,with multiple high-signal foci inside.On T2WI fat-suppression sequence,a low-signal mass was observed with multiple patchy high signals inside.Three days after admission,the patient underwent lesion resection.Intraoperatively,the tumor was located within the fascia,between the medial quadriceps muscles,presenting as lipomatous tissue with an intact capsule,mildly adherent to surrounding tissue,and with partial muscle invasion.The mass and invaded muscle were completely excised.The postoperative pathological examination results revealed a gray-yellow nodule measuring 16.0 cm.× 10.0 cm×4.0 cm,with a smooth surface and intact capsule.The cut surface was gray-yellow,fatty-like,and soft in texture.Additional gray-yellow tissue fragments were found,with a total volume of 5.0 cm×4.0 cm×1.2 cm,exhibiting a gray-brown cut surface with moderate texture.Under microscope,the lesion consisted mainly of relatively mature adipose tissue.Enlarged and hyperchromatic nuclei were observed,with scattered mononuclear or multinuclear atypical stromal cells.Fibrous septa contained variable numbers of univacuolated or multivacuolated lipoblasts.The immunohistochemistry results showed positivity for cyclin-dependent kinase 4(CDK4)and murine double minute 2(MDM2).The fluorescence in situ hybridization(FISH)results demonstrated MDM2 gene amplification in tumor cells.The pathological diagnosis was ALT of the left lower extremity.At 6-month follow-up after operation,no tumor recurrence was observed.The preoperative MRI detection may provide effective evidence for the diagnosis of ALT,while postoperative pathological examination can confirm the diagnosis and help evaluate the prognosis of the patients.

This paper presents a rare case of renal amyloidosis complicated with primary minimal change disease. The patient initially presented with edema and proteinuria, accompanied by IgG-λ monoclonal immunoglobulinemia, leading to a diagnosis of primary systemic immunoglobulin light chain amyloidosis with renal involvement. Following treatment, the patient achieved both hematologic and renal remission. However, a renal relapse occurred two years later, presenting as nephrotic syndrome without hematologic disease recurrence. A repeat renal biopsy revealed no obvious change in amyloid deposition, but demonstrated markedly enlarged effacement of podocyte foot processes. Based on these findings, a secondary diagnosis of primary minimal change disease was established. The patient exhibited a rapid response to immunosuppressive therapy, achieving sustained long-term remission. This case underscores the importance of remaining vigilant to etiological changes in the treatment of renal diseases and highlights the role of repeated renal biopsy in refining the diagnosis and guiding treatment.

Objective:To investigate the causal impact of maternal smoking around birth(MSAB)on off-spring's risk of attention deficit hyperactivity disorder(ADHD),autism spectrum disorder(ASD),bipolar disorder(BD),and major depressive disorder(MDD).Methods:The datasets for MSAB and 4 psychiatric disorders were extracted from genome-wide association studies(GWAS).Mendelian randomization(MR)was employed,using in-verse variance weighting(IVW)as the primary analysis method.Sensitivity analyses and outlier correction were conducted using weighted median(WM),MR-Egger regression,and MR-PRESSO.The results were expressed as odds ratios(OR)and corrected for false discovery rate(FDR).Results:MR analysis showed significant causal re-lationships between MSAB and increased risk of ADHD(OR=5.36,95％CI=2.58-7.63,PFDR=0.003),MDD(OR=1.92,95％CI=1.29-2.88,PFDR=0.003),and BD(OR=6.33,95％CI=1.56-8.73,PFDR=0.013).However,no statistically significant association was found between MSAB and ASD(OR=1.66,95％CI=0.23-5.87,PFDR=0.616).Conclusion:This study suggests a potential causal link between maternal smoking around the time of birth and an increased risk of attention deficit hyperactivity disorder,bipolar disorder,and major depressive disorder in offspring.

Objective To develop a graded early mobility implementation pathway for critically ill adult patients in tertiary hospitals in Beijing and to preliminarily validate its feasibility and effectiveness.Methods Based on the"goal-directed"early mobility concept,a graded early mobility implementation pathway for critically ill patients was developed through evidence synthesis and the Delphi method,consisting of 3 components:patient inclusion,mobility implementation,and mobility evaluation.Using convenience sampling,patients meeting inclusion criteria in the general ICU of a tertiary hospital in Beijing from October 2024 to January 2025 were selected as participants.Among them,25 patients admitted from December 2024 to January 2025 were assigned to an experimental group and received early mobility interventions following the developed pathway.25 patients admitted from October to November 2024 served as a control group and received routine ICU mobility care.Outcomes including diaphragm excursion,muscle strength,ICU length of stay,and adverse events were compared between the 2 groups.Results The graded early mobility pathway achieved an implementation rate of 70.05％in the experimental group,significantly higher than it in the control group(P＜0.001),without increasing adverse events.Post-intervention diaphragm excursion in the experimental group was significantly greater than that in the control group(P=0.018).Conclusion The developed graded early mobility implementation pathway for ICU patients demonstrates scientific rigor and clinical practicality.It provides a reference for the widespread and effective implementation of early mobility in ICUs,standardizing its clinical application.

Objective:To systematically evaluate the home care needs of the disabled elderly in China, to provide a reference for developing home care services oriented to the needs of the elderly.Methods:Qualitative studies on the need for home care services for the disabled elderly in China were electronically searched in PubMed, Web of Science, Embase, Cochrane Library, China National Knowledge Infrastructure, VIP, WanFang Data, and China Biology Medicine disc. The search time limit was from the tstablishment of the database to March 9, 2024. The methodological quality of the included literature was evaluated using the quality evaluation criteria for qualitative research (2016) of the Joanna Briggs Institute Evidence-Based Health Care Center. The results of the qualitative study were analyzed using the convergent integration method.Results:A total of 12 articles were included, and 43 outcomes were distilled, summarized to form eight new categories, and synthesized to produce two outcomes, including medical care and rehabilitation needs (professional nursing needs, rehabilitation and exercise needs, health guidance needs, and psychological guidance needs), and supportive needs (policy support needs, home environment needs, service resource needs, and life care needs) .Conclusions:Disabled elderly have multifaceted needs for home care. It is recommended that a multidisciplinary team be formed and a multi-pronged integrated care model be adopted to meet the needs of the disabled elderly living at home.

This paper presents a rare case of renal amyloidosis complicated with primary minimal change disease. The patient initially presented with edema and proteinuria, accompanied by IgG-λ monoclonal immunoglobulinemia, leading to a diagnosis of primary systemic immunoglobulin light chain amyloidosis with renal involvement. Following treatment, the patient achieved both hematologic and renal remission. However, a renal relapse occurred two years later, presenting as nephrotic syndrome without hematologic disease recurrence. A repeat renal biopsy revealed no obvious change in amyloid deposition, but demonstrated markedly enlarged effacement of podocyte foot processes. Based on these findings, a secondary diagnosis of primary minimal change disease was established. The patient exhibited a rapid response to immunosuppressive therapy, achieving sustained long-term remission. This case underscores the importance of remaining vigilant to etiological changes in the treatment of renal diseases and highlights the role of repeated renal biopsy in refining the diagnosis and guiding treatment.

Purpose To explore the diagnostic efficacy of texture analysis in grading multiple ligament injuries of the knee Joint.Materials and Methods MRI images of 203 patients with knee injuries in the Second Affiliated Hospital of Shantou University School of Medicine from June 2018 to September 2020 were retrospectively collected,and then graded the injuries of multiple ligaments.Extraction of texture features and discriminant analysis were accomplished by MaZda software.Spearman correlation analysis and LASSO penalized Logistic regression analysis were used to further explore the differential diagnostic efficacy of each texture parameter for grading knee ligament injuries.Results(1)MaZda software was found to have the highest diagnostic efficacy for patellar ligament injury grading by using different texture feature screening methods in combination with discriminant analysis.Non-linear discriminant analysis had the lowest misclassification rate of 4.43%.(2)The texture parameters that were ultimately included in the analysis of the subjects'work characteristic curves appeared more frequently in the gray level co-occurrence matrix and the run length matrix.(3)The area under the curve of the texture parameters finally included in the analysis were 0.870-0.975,0.772-0.912,0.836-0.929 and 0.766-0.920,respectively in patellar ligament,anterior cruciate ligament,posterior cruciate ligament and lateral collateral ligament for the diagnosis of knee ligament injury grading.The diagnostic efficacy for patellar ligament injury grading was the highest,which was consistent with the lowest misclassification rate for patellar ligament in the discriminant analysis of texture features.And the texture parameters ultimately included in patellar ligament were all highly correlated with its injury grading.Conclusion The texture analysis technique has high diagnostic efficacy for grading knee ligament injuries,and can provide a more accurate diagnostic imaging basis for the clinic.

Objective:To systematically evaluate the home care needs of the disabled elderly in China, to provide a reference for developing home care services oriented to the needs of the elderly.Methods:Qualitative studies on the need for home care services for the disabled elderly in China were electronically searched in PubMed, Web of Science, Embase, Cochrane Library, China National Knowledge Infrastructure, VIP, WanFang Data, and China Biology Medicine disc. The search time limit was from the tstablishment of the database to March 9, 2024. The methodological quality of the included literature was evaluated using the quality evaluation criteria for qualitative research (2016) of the Joanna Briggs Institute Evidence-Based Health Care Center. The results of the qualitative study were analyzed using the convergent integration method.Results:A total of 12 articles were included, and 43 outcomes were distilled, summarized to form eight new categories, and synthesized to produce two outcomes, including medical care and rehabilitation needs (professional nursing needs, rehabilitation and exercise needs, health guidance needs, and psychological guidance needs), and supportive needs (policy support needs, home environment needs, service resource needs, and life care needs) .Conclusions:Disabled elderly have multifaceted needs for home care. It is recommended that a multidisciplinary team be formed and a multi-pronged integrated care model be adopted to meet the needs of the disabled elderly living at home.

Objective:In carbon ion treatment planning of water phantom, establish a conversion factor calculation system and conversion factor curves for organs at risk (OAR) for microdosimetric kinetic models (MKM) and local effect models (LEM), and validate them in clinical patient planning.Methods:Using a uniform spherical water phantom as the research object, relative biological effectiveness-weighted doses (RWD) for the LEM were re-calculated based on the physical dose of RayStation-MKM. The median dose within the planning target volume (PTV) of LEM and MKM was regarded as the conversion factor. The impacts of single-fraction target prescription dose, spread-out Bragg peak (SOBP) width and depth, shape, and irradiation mode on the conversion factor were assessed, and a conversion factor calculation system was established. Additionally, the accuracy of the conversion factor calculation system was validated using both water phantoms and clinical patient cases. The conversion factor curves for OAR were computed based on clinical patient treatment plans.Results:The primary influencing factors for the conversion factors were the single-fraction prescription dose, target SOBP width and depth. The conversion factors were increased with the increase of SOBP width and target depth, whereas decreased with the increase of the single-fraction prescription dose. Under single-field irradiation, a conversion factor calculation system was established based on above 3 parameters. For the plans of 9 patients, the average difference between the calculated results and the conversion factor calculation system was 0.340% ± 0.203%, and the average difference in the conversion curves for OAR was 2.650% ± 2.399%.Conclusion:A dose conversion factor calculation system and conversion factor curves for OAR for carbon ion radiotherapy are established for MKM and LEM, and their accuracy meets the requirements for use in clinical patient treatment plans.