Peptide-based radiopharmaceuticals targeting integrin α5β1 show promise for precise tumor diagnosis and treatment. However, current peptide-based radioligands that target α5β1 demonstrate inadequate in vivo performance owing to limited tumor retention. The use of PEGylation to enhance the tumor retention of radiopharmaceuticals by prolonging blood circulation time poses a risk of increased blood toxicity. Therefore, a PEGylation strategy that boosts tumor retention while minimizing blood circulation time is urgently needed. Here, we developed a PEGylation-enabled peptide multidisplay platform (PEGibody) for PR_b, an α5β1 targeting peptide. PEGibody generation involved PEGylation and self-assembly. [⁶⁴Cu]QM-2303 PEGibodies displayed spherical nanoparticles ranging from 100 to 200 nm in diameter. Compared with non-PEGylated radioligands, [⁶⁴Cu]QM-2303 demonstrated enhanced tumor retention time due to increased binding affinity and stability. Importantly, the biodistribution analysis confirmed rapid clearance of [⁶⁴Cu]QM-2303 from the bloodstream. Administration of a single dose of [¹⁷⁷Lu]QM-2303 led to robust antitumor efficacy. Furthermore, [⁶⁴Cu]/[¹⁷⁷Lu]QM-2303 exhibited low hematological and organ toxicity in both healthy and tumor-bearing mice. Therefore, this study presents a PEGibody-based radiotheranostic approach that enhances tumor retention time and provides long-lasting antitumor effects without prolonging blood circulation lifetime. The PEGibody-based radiopharmaceutical [⁶⁴Cu]/[¹⁷⁷Lu]QM-2303 shows great potential for positron emission tomography imaging-guided targeted radionuclide therapy for α5β1-overexpressing tumors.

Objective: To construct a Family with sequence similarity 50 member A(FAM50A) gene knockout mouse insulinoma pancreatic β-cell line Beta-TC-6 using CRISPR/Cas9 gene editing technology and to prepare polyclonal antibodies specifically recognizing FAM50A. Methods: Two guide RNAs(sgRNAs) targeting the FAM50A gene were designed,and a recombinant plasmid expressing blue fluorescent protein(BFP) was constructed for gene knockout.The successfully constructed plasmid was transfected into Beta-TC-6 cells,and BFP-positive single cells were isolated for clonal expansion.The expanded monoclonal cell lines were genotyped by Sanger sequencing,and FAM50A protein expression was assessed by Western blot.Purified human recombinant FAM50A protein was used to immunize New Zealand rabbits for the preparation of a polyclonal antibody.The specificity of the prepared antibody was then validated using the successfully established FAM50A knockout cell line. Results: A monoclonal cell line with a successful knockout of the FAM50A gene was identified.Sanger sequencing confirmed base deletions at the target site.Western blot analysis showed a complete absence of FAM50A protein expression in this cell line.The prepared polyclonal antibody successfully recognized endogenous murine FAM50A protein in wild-type Beta-TC-6 cells and in hTERT-RPE1 cells overexpressing human FAM50A-GFP fusion protein,while no signal was detected in the FAM50A knockout cells. Conclusion This study successfully established a FAM50A gene knockout Beta-TC-6 cell model and generated a FAM50A polyclonal antibody,providing powerful tools for future research.

BACKGROUND:High-methionine diet can cause liver injury in Cbs+/-mice,and hyperhomocystinemia is related to the occurrence and progression of various liver-related diseases,such as hepatic steatosis,autoimmune hepatitis,and alcoholic fatty liver disease.MicroRNAs(miRNAs)are involved in various cellular processes including cell survival,differentiation and autophagy,which are of great significance. OBJECTIVE:To investigate the critical role of miR-144-3p on Cbs+/-mouse hepatocyte autophagy induced by high methionine die. METHODS:(1)Ten male cystathione-β-synthase normal(Cbs+/+)mice and another 10 male mice with single gene knockout(Cbs+/-)of similar body mass,4 weeks of age,were fed a high-methionine diet and executed after 12 weeks to take liver tissue.(2)Human hepatocytes(HL-7702)were cultured in vitro and divided into control[0 μmol/L homocysteine(Hcy)],Hcy(100 μmol/L Hcy),mimic-NC(transfected with mimic-NC),mimic-NC + Hcy(mimic-NC transfecton+100 μmol/L Hcy),miR-144-3p mimic(transfected with miR-144-3p mimic),and miR-144-3p mimic + Hcy(miR-144-3p mimic transfection+100 μ mol/L Hcy),inhibitor-NC(transfected with inhibitor-NC),inhibitor-NC + Hcy(inhibitor-NC transfection + 100 μmol/L Hcy),miR-144-3p inhibitor(transfected with miR-144-3p inhibitor),and miR-144-3p inhibitor + Hcy(miR-144-3p inhibitor transfection + 100 μmol/L Hcy).Quantitative real-time PCR was used to detect the expression of miR-144-3p in liver tissue and hepatocytes.After transfection of miR-144-3p mimic or inhibitor,quantitative real-time PCR and western blot were used to detect the transfection efficiency of miR-144-3p and its effect on the expression of autophagy-related proteins LC3B and p62.The levels of alanine transferase and aspartate aminotransferase in hepatocyte supernatants were determined by enzyme linked immunosorbent assay.The correlation between the expression of miR-144-3 in hepatocyte and the levels of alanine transferase and aspartate aminotransferase in hepatocyte supernatants were analyzed by Pearson correlation analysis. RESULTS AND CONCLUSION:Compared with the Cbs+/+ group and control group,the expression of miR-144-3p in the liver tissue of the Cbs+/-group and in hepatocytes of the Hcy group was decreased(P<0.01).The expression of LC3B-Ⅱ/Ⅰ was decreased in hepatocyte after transfection of miR-144-3p mimic,while the protein expression of p62 was increased(P<0.01).The opposite results were obtained after transfection of miR-144-3p inhibitor(P<0.01).Compared with the mimic-NC group,the levels of alanine transferase and aspartate aminotransferase were decreased in the miR-144-3p mimic group(P<0.01),while the opposite results were obtained in the inhibitor-NC group(P<0.01).The expression of miR-144-3p in hepatocytes was negatively correlated with the levels of alanine transferase(P<0.01,r=-0.887 6)and aspartate aminotransferase(P<0.01,r=-0.829 9)in the supernatant of hepatocytes.To conclude,Hcy promotes hepatocyte autophagy by inhibiting the expression of miR-144-3p,which subsequently aggravates liver injury.

Objective To explore the application value of static progressive stretch(SPS)combined with step-wise progressive rehabilitation nursing in perioperative rehabilitation nursing of patients with unicompartmental knee arthroplasty(UKA).Methods From January 2022 to June 2023,a total of 74 patients who were scheduled to undergo UKA in the Department of Orthopedics of a tertiary A hospital in Beijing were selected by the convenient sampling method and divided into 2 groups by the random number table method,with 37 cases in each group.The experimental group was given step-by-step rehabilitation nursing combined with SPS technology,while the control group was only given step-by-step rehabilitation nursing,and the intervention lasted for 4 weeks.The knee joint activity,knee joint function score,comfort,and rehabilitation self-efficacy of the 2 groups were observed and measured before and after the intervention.Results Finally,35 patients were included in the experimental group and 36 patients in the control group.After the intervention,the knee joint range of motion,knee joint function score,comfort,rehabilitation self-efficacy and all dimensions and their total scores in the experimental group were higher than those before the intervention and those in the control group(P＜0.05).Conclusion SPS combined with progressive rehabilitation nursing can effectively improve the knee joint function and range of motion of patients after UKA,improve the comfort of patients,improve their quality of life,and enhance their rehabilitation self-efficacy,which is helpful to achieve early recovery.

Primary prostate signet ring cell carcinoma (PPSRCC) is one of the extremely rare malignant tumors in the male urogenital system, and its diagnosis mainly relies on pathological and immunohistochemical examination. Compared with typical prostate cancer, PPSRCC is characterized by more aggressive with less treatment response and poor prognosis. Current researches on the pathogenesis, clinical manifestations, pathological characteristics, diagnosis, treatment and prognosis of PPSRCC were reviewed.

By analyzing the of genetic testing data of patients with renal polycystic kidney disease and their relatives, this study aims to identify unreported novel gene mutation sites associated with autosomal dominant polycystic kidney disease (ADPKD). Structural prediction software was employed to investigate protein structural changes before and after mutations, explore genotype-phenotype correlations, and enrich the ADPKD gene database. In this single-center retrospective study, patients with multiple renal cysts diagnosed from January 2019 to February 2023 at the Zhong Da Hospital Southeast University were included. Genetic and clinical data of patients and their families were collected. Unreported novel gene mutation sites associated with ADPKD were identified. The AlphaFold v2.3.1 software was used to predict protein structures. Changes in protein structure before and after mutations were compared to explore genotype-phenotype correlations and enrich the ADPKD gene database. Twelve mutated genes associated with renal cysts were detected in 52 families. Nineteen novel gene mutation sites associated with ADPKD were identified, including 17 mutations in the PKD1 gene (one splicing mutation, seven frameshift mutations, four nonsense mutations, one whole-codon insertion, and four missense mutations); one ALG9 missense mutation; and one chromosomal structural variation. Truncating mutations in the PKD1 gene were correlated with a more severe clinical phenotype, while non-truncating mutations were associated with greater clinical heterogeneity. Numerous novel gene mutation sites associated with ADPKD remain unreported. Therefore, it is essential to analyze the pathogenicity of these novel mutation sites, establish genotype-phenotype correlations, and enrich the ADPKD gene database.

Brain organoids are self-organized 3D aggregates generated by human embryonic stem cells or human induced pluripotent stem cells. Their cell type and structure are similar to embryonic human brain, which are good in vitro models for the study of neurogenetic diseases and have been widely used in the study of neurogenetic diseases. This paper will discuss the advantages and challenges of brain organoids in the modeling of genetic diseases of the nervous system.

Objective A HPLC-quantitative analysis of multi-components by single marker(QAMS)was established to determine 5 ingredients including uracil,uridine,hypoxanthine,inosine and guanosine in Periplaneta americana.Methods Separation took place on a Agilent ZORBAX SB-Aq column(250 mm×4.6 mm,5 μm)by gradient elution of methanol-0.01 mol·L-1 potassium dihydrogen phosphate at 20℃with a flow rate of 1.0 mL·min-1.The detection wavelength was 260 nm and the injection amount was 10 μL.The relative correction factors(fa/b)was calculated for the other four components with uridine as an internal standard.The content of 5 ingredients in 10 batches of Periplaneta americana was determined by QAMS.Results were compared with those of external standard method(ESM).Results Five nucleosides showed good linear relationships in their own ranges(r＞0.999 5),and the average recoveries ranged from 97.0%to 100.8%.The relative correction factors of uracil,hypoxanthine,inosine and guanosine were 0.908 0,1.005 3,1.969 5 and 1.303 4,respectively.Conclusion The established method is accurate and stable.It can provide theoretical reference for the quality control of Periplaneta americana.

In recent years,challenges persist in the grassroots Party building in the university-affiliated hospitals though increased efforts have been intensified.These challenges include a lack of alignment between Party building and the hospital's central tasks,an absence of distinctive Party building brands,an insufficiently prominent role of Party organizations in hospital development,and inadequate leveraging of the Party construction for promoting medical services and cultivating medical talents.Starting from an exploration of the connotation and significance of constructing framework of"One Integration and Two Highs"in university-affiliated hospitals,this paper conducts an in-depth analysis of the existing problems and their causes within this frame-work.From a holistic perspective,the paper proposes a new path for establishing the"One Integration and Two Highs"frame-work in university-affiliated hospitals,centered around"Seven Integrations":concept integration,organization integration,strength integration,vehicle integration,culture integration,innovation integration,and assessment integration.

ObjectiveTo provide reference for further strengthening the management of laboratory animals in Sichuan Province by reviewing and analyzing the results of parasitic and microbial quality inspections of laboratory animals from 2017 to 2023. Methods Sichuan Province has 31 licensed laboratory animal production units, with the main species including mice, rats, guinea pigs, rabbits, dogs, monkeys, and pigs. Sampling inspections and reports were conducted for units with laboratory animal production qualifications in accordance with current national and local standards for laboratory animal classification. The quality inspection results for various laboratory animals in Sichuan Province from 2017 to 2023 were analyzed. Results With the exception of 2018, annual quality inspections of laboratory animals were conducted every year between 2017 and 2023. Mice: In 2017, positive results for helminths, flagellates, Salmonella, murine hepatitis virus antibodies, and murine pneumonia virus antibodies were detected, with a pass rate of only 42.9%. In 2019, Staphylococcus aureus and Klebsiella pneumoniae were detected, with a pass rate of 86.7%. In 2021, Sendai virus antibodies were detected, yielding a pass rate of 85.7%.The pass rate in 2020, 2022, and 2023 was 100%. Rats: In 2017, positive results were found for helminths, mycoplasma antibodies, Staphylococcus aureus, Sendai virus antibodies, murine pneumonia virus antibodies, rat parvovirus RV strain antibodies, rat parvovirus (H-1) strain antibodies, and rat coronavirus antibodies, with a pass rate of 40.0%. In 2019, mycoplasma antibodies, Staphylococcus aureus and Klebsiella pneumoniae were detected, with a pass rate of 35.0%. No positive indicators were detected in 2020. In 2021, Sendai virus antibodies and rat parvovirus RV strain antibodies were detected, with a pass rate of 50.0%. In 2022, positive results for rat parvovirus RV strain antibodies were found, yielding a pass rate of 87.5%. In 2023, Pasteurella pneumotropica and Klebsiella pneumoniae were detected, with a pass rate of 85.7%. Dogs: In 2017 and 2019, the antibody titers for rabies virus and canine distemper virus were below the required standard, with pass rates of 33.3% and 20.0%, respectively. In 2020 and 2022, the pass rate was 100%. Guinea pigs (general grade): In 2019, positive results for Toxoplasma antibodies were detected, with a pass rate of 80.0%. In all other years, the pass rate was 100%. Monkeys: In 2019, positive results for Toxoplasma gondii and rhesus herpesvirus type I antibodies were found, with a pass rate of 87.5%.In 2020 and 2022, rhesus herpesvirus type I antibodies were positively detected, yielding pass rates of 93.3% and 97.5%, respectively. The pass rates in 2021 and 2023 were 100%. Clean guinea pigs, rabbits and pigs all passed the inspection each year. ConclusionIssues related to the parasitic and microbial quality of laboratory animals persist in Sichuan Province. Supervision and sampling inspections have proven to be effective in identifying these issues promptly, serving as a critical measure to ensure the quality of laboratory animals. The results of these inspections offer valuable data to support the healthy development of the laboratory animal industry in Sichuan Province.