Gene synthesis is an enabling technology that supports the development of synthetic biology. The existing approaches for de novo gene synthesis generally have tedious operation, low efficiency, high error rates, and limited product lengths, being difficult to support the huge demand of synthetic biology. The assembly and error correction are the keys in gene synthesis. This study first designed the oligonucleotide sequences by reasonably splitting the virus genome of approximately 10 kb by balancing the parameters of sequence design software ability, PCR amplification ability, and assembly enzyme assembly ability. Then, two-step PCR was performed with high-fidelity polymerase to complete the de novo synthesis of 3.0 kb DNA fragments, and error correction reactions were performed with T7 endonuclease Ⅰ for the products from different stages of PCR. Finally, the virus genome was assembled by 3.0 kb DNA fragments from de novo synthesis and error correction and then sequenced. The experimental results showed that the proposed method successfully produced the DNA fragment of about 10 kb and reduced the probability of large fragment mutations during the assembly process, with the lowest error rate reaching 0.36 errors/kb. In summary, this study developed an efficient de novo method for synthesizing a viral genome of about 10 kb with T7 endonuclease Ⅰ-mediated error correction. This method enabled the synthesis of a 10 kb viral genome in one day and the correct plasmid of the viral genome in five days. This study optimized the de novo gene synthesis process, reduced the error rate, simplified the synthesis and assembly steps, and reduced the cost of viral genome assembly.
Genome, Viral/genetics* ; Polymerase Chain Reaction/methods* ; DNA, Viral/genetics* ; Bacteriophage T7/enzymology* ; Synthetic Biology/methods*

Peptide receptor radionuclide therapy (PRRT) with radiolabeled SSTR2 agonists is a treatment option that is highly effective in controlling metastatic and progressive neuroendocrine tumors (NETs). Previous studies have shown that an SSTR2 agonist combined with albumin binding moiety Evans blue (denoted as ¹⁷⁷Lu-EB-TATE) is characterized by a higher tumor uptake and residence time in preclinical models and in patients with metastatic NETs. This study aimed to enhance the in vivo stability, pharmacokinetics, and pharmacodynamics of ¹⁷⁷Lu-EB-TATE by replacing the maleimide-thiol group with a polyethylene glycol chain, resulting in a novel EB conjugated SSTR2-targeting radiopharmaceutical, ¹⁷⁷Lu-LNC1010, for PRRT. In preclinical studies, ¹⁷⁷Lu-LNC1010 exhibited good stability and SSTR2-binding affinity in AR42J tumor cells and enhanced uptake and prolonged retention in AR42J tumor xenografts. Thereafter, we presented the first-in-human dose escalation study of ¹⁷⁷Lu-LNC1010 in patients with advanced/metastatic NETs. ¹⁷⁷Lu-LNC1010 was well-tolerated by all patients, with minor adverse effects, and exhibited significant uptake and prolonged retention in tumor lesions, with higher tumor radiation doses than those of ¹⁷⁷Lu-EB-TATE. Preliminary PRRT efficacy results showed an 83% disease control rate and a 42% overall response rate after two ¹⁷⁷Lu-LNC1010 treatment cycles. These encouraging findings warrant further investigations through multicenter, prospective, and randomized controlled trials.

Objective:To analyze the pathogen spectrum and epidemiological characteristics of acute respiratory infections in Yantai from 2024 to 2025.Methods:Influenza-like illness（ILI）and severe acute respiratory infection（SARI）patients were enrolled from the national sentinel hospitals for acute respiratory infectious diseases in Yantai between January 2024 and April 2025. Oropharyngeal swab samples were collected and 16 common respiratory pathogens were tested using real-time quantitative PCR. Descriptive epidemiological methods were used to analyze the detection results.Results:A total of 1 499 cases were included in the study，including 1 070 ILI cases and 429 SARI cases. The overall positive rate of respiratory pathogens was 52.57%（788/1 499）. All of the 16 targeted respiratory pathogens were detected，with influenza virus A（FluA），SARS-CoV-2， Haemophilus influenzae（ Hi），and human adenovirus（HAdV）being the predominant pathogens. The co-infection rate was 6.27%（94/1 499），with FluA combined with Hi infection being the predominant type. Statistically significant differences in the positive rates of influenza B virus and SARS-CoV-2 were observed between male and female patients（both P<0.05）. The overall positive rates varied significantly across different age groups（ P<0.001），with the highest rate（64.62%，84/130）observed in the 4-14 year age group. The overall positive rates also varied significantly among different months（ P<0.001），with the highest rate in December（69.93%，107/153）. The overall positive rate was 57.48%（615/1 070）in ILI cases and 40.33%（173/429）in SARI cases，with statistically significant difference between the two groups（ P<0.001）. Influenza viruses and SARS-CoV-2 were the predominant pathogens in ILI cases，whereas HAdV and Mycoplasma pneumoniae were more common in SARI cases. Conclusions:FluA，SARS-CoV-2， Hi，and HAdV are the predominant pathogens causing acute respiratory infections in Yantai from January 2024 to April 2025. The positive rates of respiratory pathogens varies across different gender and age groups，and shows distinct seasonal patterns. There are significant differences in pathogen spectra between ILI and SARI cases.

Objective:To analyze the pathogen spectrum and epidemiological characteristics of acute respiratory infections in Yantai from 2024 to 2025.Methods:Influenza-like illness（ILI）and severe acute respiratory infection（SARI）patients were enrolled from the national sentinel hospitals for acute respiratory infectious diseases in Yantai between January 2024 and April 2025. Oropharyngeal swab samples were collected and 16 common respiratory pathogens were tested using real-time quantitative PCR. Descriptive epidemiological methods were used to analyze the detection results.Results:A total of 1 499 cases were included in the study，including 1 070 ILI cases and 429 SARI cases. The overall positive rate of respiratory pathogens was 52.57%（788/1 499）. All of the 16 targeted respiratory pathogens were detected，with influenza virus A（FluA），SARS-CoV-2， Haemophilus influenzae（ Hi），and human adenovirus（HAdV）being the predominant pathogens. The co-infection rate was 6.27%（94/1 499），with FluA combined with Hi infection being the predominant type. Statistically significant differences in the positive rates of influenza B virus and SARS-CoV-2 were observed between male and female patients（both P<0.05）. The overall positive rates varied significantly across different age groups（ P<0.001），with the highest rate（64.62%，84/130）observed in the 4-14 year age group. The overall positive rates also varied significantly among different months（ P<0.001），with the highest rate in December（69.93%，107/153）. The overall positive rate was 57.48%（615/1 070）in ILI cases and 40.33%（173/429）in SARI cases，with statistically significant difference between the two groups（ P<0.001）. Influenza viruses and SARS-CoV-2 were the predominant pathogens in ILI cases，whereas HAdV and Mycoplasma pneumoniae were more common in SARI cases. Conclusions:FluA，SARS-CoV-2， Hi，and HAdV are the predominant pathogens causing acute respiratory infections in Yantai from January 2024 to April 2025. The positive rates of respiratory pathogens varies across different gender and age groups，and shows distinct seasonal patterns. There are significant differences in pathogen spectra between ILI and SARI cases.

Calcium-based biomaterials have been intensively studied in the field of drug delivery owing to their excellent biocompatibility and biodegradability. Calcium-based materials can also deliver contrast agents, which can enhance real-time imaging and exert a Ca²⁺-interfering therapeutic effect. Based on these characteristics, amorphous calcium carbonate (ACC), as a brunch of calcium-based biomaterials, has the potential to become a widely used biomaterial. Highly functional ACC can be either discovered in natural organisms or obtained by chemical synthesis However, the standalone presence of ACC is unstable in vivo. Additives are required to be used as stabilizers or core-shell structures formed by permeable layers or lipids with modified molecules constructed to maintain the stability of ACC until the ACC carrier reaches its destination. ACC has high chemical instability and can produce biocompatible products when exposed to an acidic condition in vivo, such as Ca²⁺ with an immune-regulating ability and CO₂ with an imaging-enhancing ability. Owing to these characteristics, ACC has been studied for self-sacrificing templates of carrier construction, targeted delivery of oncology drugs, immunomodulation, tumor imaging, tissue engineering, and calcium supplementation. Emphasis in this paper has been placed on the origin, structural features, and multiple applications of ACC. Meanwhile, ACC faces many challenges in clinical translation, and long-term basic research is required to overcome these challenges. We hope that this study will contribute to future innovative research on ACC.

Objective To explore the potential pathogenic genes of intestinal metaplasia.Methods Twenty-one patients with intestinal metaplasia admitted to the Department of Gastroenterology at the Second Affiliated Hospital of Anhui University of Chinese Medicine from January,2022 to June,2022,and 21 healthy subjects undergoing gastroscopic examination during the same period were enrolled in this study.All the participants underwent gastroscopy and pathological examination,and gastric tissue samples were collected for transcriptome sequencing to screen for differentially expressed genes(DEGs).The biological functions of the DEGs were analyzed using bioinformatics analysis,and qRT-PCR was used to validate the results.Results Transcriptomic sequencing identified a total of 1373 DEGs,including 827 upregulated and 546 downregulated ones.The top 6 upregulated genes(AGMAT,CCL25,FABP1,CDX1,SPINK4,and MUC2),ranked based on their significance and average expression level,were selected for validation,and qRT-PCR showed significant upregulation of their mRNAs in the gastric tissues of patients with intestinal metaplasia(P<0.05).Conclusion AGMAT,CCL25,FABP1,CDX1,SPINK4,and MUC2 participate in the occurrence and development of intestinal metaplasia,and may serve as potential biomarkers for diagnosing intestinal metaplasia.

Objective To explore the potential pathogenic genes of intestinal metaplasia.Methods Twenty-one patients with intestinal metaplasia admitted to the Department of Gastroenterology at the Second Affiliated Hospital of Anhui University of Chinese Medicine from January,2022 to June,2022,and 21 healthy subjects undergoing gastroscopic examination during the same period were enrolled in this study.All the participants underwent gastroscopy and pathological examination,and gastric tissue samples were collected for transcriptome sequencing to screen for differentially expressed genes(DEGs).The biological functions of the DEGs were analyzed using bioinformatics analysis,and qRT-PCR was used to validate the results.Results Transcriptomic sequencing identified a total of 1373 DEGs,including 827 upregulated and 546 downregulated ones.The top 6 upregulated genes(AGMAT,CCL25,FABP1,CDX1,SPINK4,and MUC2),ranked based on their significance and average expression level,were selected for validation,and qRT-PCR showed significant upregulation of their mRNAs in the gastric tissues of patients with intestinal metaplasia(P<0.05).Conclusion AGMAT,CCL25,FABP1,CDX1,SPINK4,and MUC2 participate in the occurrence and development of intestinal metaplasia,and may serve as potential biomarkers for diagnosing intestinal metaplasia.

Objective To monitor the susceptibility of clinical isolates to antimicrobial agents in healthcare facilities in major regions of China in 2023.Methods Clinical isolates collected from 73 hospitals across China were tested for antimicrobial susceptibility using a unified protocol based on disc diffusion method or automated testing systems.Results were interpreted using the 2023 Clinical & Laboratory Standards Institute (CLSI) breakpoints.Results A total of 445199 clinical isolates were collected in 2023,of which 29.0％ were gram-positive and 71.0％ were gram-negative.The prevalence of methicillin-resistant strains in Staphylococcus aureus,Staphylococcus epidermidis and other coagulase-negative Staphylococcus species (excluding Staphylococcus pseudintermedius and Staphylococcus schleiferi) (MRSA,MRSE and MRCNS) was 29.6％,81.9％ and 78.5％,respectively.Methicillin-resistant strains showed significantly higher resistance rates to most antimicrobial agents than methicillin-susceptible strains (MSSA,MSSE and MSCNS).Overall,92.9％ of MRSA strains were susceptible to trimethoprim-sulfamethoxazole and 91.4％ of MRSE strains were susceptible to rifampicin.No vancomycin-resistant strains were found.Enterococcus faecalis had significantly lower resistance rates to most antimicrobial agents tested than Enterococcus faecium.A few vancomycin-resistant strains were identified in both E.faecalis and E.faecium.The prevalence of penicillin-susceptible Streptococcus pneumoniae was 93.1％ in the isolates from children and and 95.9％ in the isolates from adults.The resistance rate to carbapenems was lower than 15.0％ for most Enterobacterales species except for Klebsiella,22.5％ and 23.6％ of which were resistant to imipenem and meropenem,respectively .Most Enterobacterales isolates were highly susceptible to tigecycline,colistin and polymyxin B,with resistance rates ranging from 0.6％ to 10.0％.The resistance rate to imipenem and meropenem was 21.9％ and 17.4％ for Pseudomonas aeruginosa,respectively,and 67.5％ and 68.1％ for Acinetobacter baumannii,respectively.Conclusions Increasing resistance to the commonly used antimicrobial agents is still observed in clinical bacterial isolates.However,the prevalence of important crabapenem-resistant organisms such as crabapenem-resistant K.pneumoniae,P.aeruginosa,and A.baumannii showed a slightly decreasing trend.This finding suggests that strengthening bacterial resistance surveillance and multidisciplinary linkage are important for preventing the occurrence and development of bacterial resistance.

Objective To monitor the susceptibility of clinical isolates to antimicrobial agents in healthcare facilities in major regions of China in 2023.Methods Clinical isolates collected from 73 hospitals across China were tested for antimicrobial susceptibility using a unified protocol based on disc diffusion method or automated testing systems.Results were interpreted using the 2023 Clinical & Laboratory Standards Institute (CLSI) breakpoints.Results A total of 445199 clinical isolates were collected in 2023,of which 29.0％ were gram-positive and 71.0％ were gram-negative.The prevalence of methicillin-resistant strains in Staphylococcus aureus,Staphylococcus epidermidis and other coagulase-negative Staphylococcus species (excluding Staphylococcus pseudintermedius and Staphylococcus schleiferi) (MRSA,MRSE and MRCNS) was 29.6％,81.9％ and 78.5％,respectively.Methicillin-resistant strains showed significantly higher resistance rates to most antimicrobial agents than methicillin-susceptible strains (MSSA,MSSE and MSCNS).Overall,92.9％ of MRSA strains were susceptible to trimethoprim-sulfamethoxazole and 91.4％ of MRSE strains were susceptible to rifampicin.No vancomycin-resistant strains were found.Enterococcus faecalis had significantly lower resistance rates to most antimicrobial agents tested than Enterococcus faecium.A few vancomycin-resistant strains were identified in both E.faecalis and E.faecium.The prevalence of penicillin-susceptible Streptococcus pneumoniae was 93.1％ in the isolates from children and and 95.9％ in the isolates from adults.The resistance rate to carbapenems was lower than 15.0％ for most Enterobacterales species except for Klebsiella,22.5％ and 23.6％ of which were resistant to imipenem and meropenem,respectively .Most Enterobacterales isolates were highly susceptible to tigecycline,colistin and polymyxin B,with resistance rates ranging from 0.6％ to 10.0％.The resistance rate to imipenem and meropenem was 21.9％ and 17.4％ for Pseudomonas aeruginosa,respectively,and 67.5％ and 68.1％ for Acinetobacter baumannii,respectively.Conclusions Increasing resistance to the commonly used antimicrobial agents is still observed in clinical bacterial isolates.However,the prevalence of important crabapenem-resistant organisms such as crabapenem-resistant K.pneumoniae,P.aeruginosa,and A.baumannii showed a slightly decreasing trend.This finding suggests that strengthening bacterial resistance surveillance and multidisciplinary linkage are important for preventing the occurrence and development of bacterial resistance.

Objective:To investigate the serum levels of soluble low-density lipoprotein receptor 11 (sLR11), endothelial cell-specific molecule-1 (ESM-1), and advanced glycosylation end products (AGE) in patients with preeclampsia (PE) and their ability to predict adverse pregnancy outcomes.Methods:A total of 141 PE patients (PE group) and 60 normal pregnant women (control group) who were admitted to the Zhangjiakou Maternal and Child Health Hospital from January 2020 to October 2022 were selected. Serum levels of sLR11, ESM-1, and AGE were detected in each group. PE patients were divided into mild preeclampsia (MP, n=78) and severe preeclampsia (SP, n=63) according to the severity of the disease. PE patients were also divided into an adverse pregnancy outcome group ( n=57) and a good pregnancy outcome group ( n=84) based on the occurrence of adverse pregnancy outcomes. Receiver operating characteristic (ROC) curve analysis was used to evaluate the ability of serum levels of sLR11, ESM-1, and AGE to predict adverse pregnancy outcomes in PE patients. Pearson correlation analysis was used to examine the correlation between serum levels of sLR11, ESM-1, and AGE in PE patients. Results:Serum levels of sLR11, ESM-1, and AGE were significantly higher in the PE group than in the control group (all P<0.001). Serum levels of sLR11, ESM-1, and AGE were significantly higher in the SP group than in the MP group (all P<0.001). Serum levels of sLR11, ESM-1, and AGE were significantly higher in the adverse pregnancy outcome group than in the good pregnancy outcome group (all P<0.001). ROC curve analysis showed that the combination of sLR11≥11.65 μg/L, ESM-1≥2.14 μg/L, and AGE≥57.38 ng/ml had the largest area under the curve (AUC) for predicting adverse pregnancy outcomes in PE patients (0.947, 95% CI: 0.890-0.995), with a sensitivity of 97.3% and specificity of 82.0%. Pearson correlation analysis showed that serum levels of sLR11, ESM-1, and AGE were positively correlated in PE patients (all P<0.001). Conclusions:Serum levels of sLR11, ESM-1, and AGE are significantly increased in PE patients and are closely related to disease severity. The combination of these three factors has good value in predicting adverse pregnancy outcomes in PE patients.