Objective Aiming at detecting Staphylococcus aureus、Pseudomonas aeruginosa and Klebsiella pneumoniae in laboratory animals,the paper provides a rapid,sensitive and simple test method.Methods According to Staphylococcus aureus nuc gene,Pseudomonas aeruginosa LasI gene,Klebsiella pneumonia PhoE gene and general 16S rRNA gene, designed specific primers;Through the optimization of multiplex PCR primer concentrations and annealing temperature, the specificity and sensitivity of detection, establishing multiplex PCR system.Application of the PCR system test specimens of artificial infections and experiment animal feces is compared with traditional test method.Results Multiplex PCR amplification of Staphylococcus aureus (153 bp), Pseudomonas aeruginosa (600 bp) with Klebsiella pneumoniae (368 bp) and general (520 bp).The multiplex sensitivity for the purpose of 10pg, specificity of detection was not detected from other pathogens.Application of establishing multiplex PCR system to detect the artificial positive samples, and detect 1 Pseudomonas aeruginosa positive case in 76 fecals.Conclusions This paper established the multiplex PCR method which has the advantages of specific,sensitive,simple and rapid, and provides a reliable way for rapid test in laboratory animals microbiology.

The aim of this study is to establish a multiplex polymerase chain raction (PCR) to identify of four kinds of laboratory animal pathogens: Pasteurella multocida, Bordetella bronchiseptica, Mycoplasma pneumoniae and Klebsiella pneumoniae.Methods Specific primers were designed based on GenBank data.The multiplex PCR system was established through optimization of multiple PCR and detection of its specificity and sensitivity.This technique was used to test artificially infected samples and tracheal secretions of experimental animals (rat, mouse, guinea pig, rabbit, hamster), and comparing the detection results by this method and traditional detection test.Results Target bands of Pasteurella multocida (356 bp), Bordetella bronchiseptica (237 bp), Mycoplasma pneumoniae (266 bp), and Klebsiella pneumoniae (142 bp) were obtained, with a detection sensitivity of Klebsiella pneumoniae of 10 pg, and that of Pasteurella multocida, Bordetella bronchiseptica and Mycoplasma pneumoniae of 1 pg by this newly developed multiplex PCR assay.No target bands were observed from the non-specific pathogens of artificially infected samples.The tracheal secretions taken from 45 experimental animals (mice and rabbits) were tested with this new PCR assay, among which 15 cases of Klebsiella pneumonia and 9 cases of Pasteurella multocida were detected as positive, while all the results of traditional method and serological test were negative.Conclusions A simple, rapid, specific and highly sensitive multiplex PCR system has been successfully established.It is valuable for detection of Pasteurella multocida, Bordetella bronchiseptica, Mycoplasma pneumoniae, and Klebsiella pneumoniae in laboratory animals.

Chuangxinmycin (CM) from Actinoplanes tsinanensis was an antibiotic discovered by Chinese scientists about 40 years ago. It contains a new heterocyclic system of indole fused with dihydrothiopyran, whose biosynthetic mechanism remains unclear. CM is used as an oral medicine in the treatment of bacterial infections in China. The simple structure makes CM as an attractive candidate of structure modification for improvement of antibacterial activity. Recently, we analyzed the secondary metabolites of Actinoplanes tsinanensis CPCC 200056, a CM producing strain, as a natural CM analogue. We discovered the first natural CM analogue 3-demethylchuangxinmycin (DCM) as a new natural product. Compared to CM, DCM exhibited a much weaker activity in the inhibition of the bacterial strains tested. The finding provides valuable information for the structure-activity relationship in the biosynthesis of CM.

In order to prepare monoclonal antibody to σ A protein of avian reovirus(ARV)and es-tablish a sandwich ELISA method for the detection of ARV pathogens.In this study,the σ A pro-tein of ARV was expressed as antigen by prokaryotic expression and used to immunize BALB/c mice.Then,stable hybridoma cell lines were screened,and monoclonal antibodies were prepared.A sandwich ELISA detection method based on monoclonal antibody of σA protein was established,and the sensitivity,specificity,repeatability,and accuracy were tested.The results showed that the recombinant plasmid pET-32a-σA was successfully constructed and well expressed in Escherichia coli.After immunizing mice,two hybridoma cell lines 6B3 and 8E11,which could secrete mono-clonal antibodies stably,were successfully prepared.Both monoclonal antibodies could react with natural ARV.One of the monoclonal antibodies secreted by 6B3 was selected as the capture anti-body and the ARV-positive chicken polyclonal antibody was used as the detection antibody.A sand-wich ELISA method was established to detect ARV by optimizing the reaction conditions.The specific test showed that the method only detected ARV pathogens and no other common chicken viral pathogens were detected.The detection limit was 7.72 X 102 EID50/mL of ARV antigen.The coefficient of variation of the intra-and inter-assay tests were less than 5.0％and the reproducibili-ty was good.Thirty samples were tested simultaneously by σA-sandwich ELISA and PCR,and the results were consistent with each other.In conclusion,a sandwich ELISA method based on the monoclonal antibody of σA protein was successfully established for the identification and detection of ARV,which provided a technical means for the accurate and rapid detection of ARV.

OBJECTIVE To know about the development trend of small nucleic acid drugs in the world ，to provide reference for the research and development of small nucleic acid drug in China. METHODS By searching the academic literature and patents related to small nucleic acid drugs through the Web of Science literature database and PatSnap patent database from Jan. 1980 to Dec. 2021，research and development situation of small nucleic acid drugs were revealed comprehensively by analyzing research enthusiasm，R&D countries ，R&D institutions and technical topics of small nucleic acid drugs. RESULTS & CONCLUSIONS A total of 59 819 documents and 37 645 patent groups were included. The global trend of small nucleic acid drug literature publication and patent application could be divided into three stages. From 2003 to 2021，the research enthusiasm for small nucleic acid drugs continued to increase. The United States ，China，Japan and Germany were the main research and development countries for small nucleic acid drugs. The number of document publications （25 703，15 927 papers）and patent applications （14 240、8 937 groups） in the United States and China were ahead of other countries ，and the research and development activities were relatively strong. Moreover，the number of document publications and patent applications in China in this field had grown rapidly in recent years. The R&D institution with the largest number of publications was the University of California （2 499 papers），the R&D institution with the largest number of patent applications was the American Ionis Corporation （1 378 groups），and the Chinese Academy of Sciences （1 580 papers）had been shortlisted among the top 10 document producing institutions in the world. However ，our country ’s research and development in this field are mostly based on basic research ，and the research on industrial application is slightly insufficient. The research focus in the field of small nucleic acid drugs mainly focuses on nucleic acid sequences and their modification and improvement and drug loading technology. RNA interference technology has gradually become a hot technology for small nucleic acid drugs.

【Objective】 To analyze nutrition-related factors that affect the prevalence of myopia in adolescents, in order to provide reference for primary prevention of myopia. 【Methods】 A stratified sampling method was used to select 385 adolescents from Zhengding County in October 2021. Adolescents in this study took vision testing, physical examination and completed a questionnaire survey. One-way analysis of variance and lasso regression were used to screen the variables, and Logistic regression was used to determine the possible influencing factors of myopia. Factor analysis was adopted to extract the dietary patterns of adolescents in Zhengding County, then the association between dietary patterns and myopia was analyzed. 【Results】 A total of 385 adolescents were surveyed, with the prevalence rate of myopia of about 68.6%. Multivariate analysis revealed that fried food(OR=8.480, 95%CI:1.058 - 67.971) was a risk factor for myopia, while intake of milk and dairy products(OR=0.994, 95%CI:0.991 - 0.999), soybeans and nuts(OR=0.997, 95%CI:0.994 - 0.999), no myopia in either parent(OR=0.312, 95%CI:0.115 - 0.845), physical education class 4 times per week(OR=0.269, 95%CI:0.074 - 0.984) were statistically associated with a lower risk of myopia(P<0.05). Three dietary patterns was extracted through factor analysis, including diversified dietary pattern, soy-hybrid dietary pattern, and snack and beverage dietary pattern. Logistic analysis results indicated that soy-hybrid dietary pattern(OR=0.85, 95%CI:0.73 - 0.99,P<0.05) was statistically associated with myopia. 【Conclusions】 The problem of myopia among adolescents in Zhengding County is more serious. Increasing the intake of milk and dairy products, soybeans and nuts, reducing the intake of fried foods, and adjusting the overall dietary structure should be recommended in order to prevent the development of myopia in adolescents.

OBJECTIVE To analyze technical and legal information of patents in the field of antigen immunoassay technology, and to provide advice to related enterprises on R&D direction and patent layout. METHODS Using patent analysis means combined with data visualization presentation, the R&D trends and research hotspots in the field of immunoassays of Abbott, Wondfo Bio, BGI Genomics and other related companies were systematically analyzed. RESULTS Abbott's patent layout in the field of immunoassay was more comprehensive and occupied a dominant position in international competition. Although the immunoassay technologies of Chinese enterprises had their own expertise, the industrial layout and the comprehensiveness of patent layout were weak. CONCLUSION Chinese enterprises should strengthen interdisciplinary research and strive for breakthroughs in the direction of cutting-edge technologies such as multiplex multi-pathogen high-throughput detection, magnetic particle chemiluminescence detection and microfluidic chip detection, as well as choosing appropriate patent protection strategies considering their own characteristics.

BACKGROUND: In China, lung cancer remains the cancer with the highest incidence and mortality rate. Among early-stage lung adenocarcinomas (LUAD), the micropapillary (MPP) component is prevalent and typically exhibits high aggressiveness, significantly correlating with early metastasis, lymphatic infiltration, and reduced five-year survival rates. Therefore, the study is to explore the similarities and differences between MPP and non-micropapillary (non-MPP) components in malignant pulmonary nodules characterized by GGOs in early-stage LUAD, identify unique mutational features of the MPP component and analyze the relationship between the ZNF469 gene, a member of the zinc-finger protein family, and the prognosis of early-stage LUAD, as well as its correlation with immune infiltration. METHODS: A total of 31 malignant pulmonary nodules of LUAD were collected and dissected into paired MPP and non-MPP components using microdissection. Whole-exome sequencing (WES) was performed on the components of early-stage malignant pulmonary nodules. Mutational signatures analysis was conducted using R packages such as maftools, Nonnegative Matrix Factorization (NMF), and Sigminer to unveil the genomic mutational characteristics unique to MPP components in invasive LUAD compared to other tumor tissues. Furthermore, we explored the expression of the ZNF469 gene in LUAD using The Cancer Genome Atlas (TCGA) database to investigate its potential association with the prognosis. We also investigated gene interaction networks and signaling pathways related to ZNF469 in LUAD using the GeneMANIA database and conducted Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. Lastly, we analyzed the correlation between ZNF469 gene expression and levels of immune cell infiltration in LUAD using the TIMER and TISIDB databases. RESULTS: MPP components exhibited a higher number of genomic variations, particularly the 13th COSMIC (Catalogue of Somatic Mutations in Cancer) mutational signature characterized by the activity of the cytidine deaminase APOBEC family, which was unique to MPP components compared to non-MPP components in tumor tissues. This suggests the potential involvement of APOBEC in the progression of MPP components in early-stage LUAD. Additionally, MPP samples with high similarity to APOBEC signature displayed a higher tumor mutational burden (TMB), indicating that these patients may be more likely to benefit from immunotherapy. The expression of ZNF469 was significantly upregulated in LUAD compared to normal tissue, and was associated with poor prognosis in LUAD patients (P<0.05). Gene interaction network analysis and GO/KEGG enrichment analysis revealed that COL6A1, COL1A1, COL1A2, TGFB2, MMP2, COL8A2 and C2CD4C interacted with ZNF469 and were mainly involved in encoding collagen proteins and participating in the constitution of extracellular matrix. ZNF469 expression was positively correlated with immune cell infiltration in LUAD (P<0.05). CONCLUSIONS The study has unveiled distinctive mutational signatures in the MPP components of early-stage invasive LUAD in the Asian population. Furthermore, we have identified that the elevated expression of mutated ZNF469 impacts the prognosis and immune infiltration in LUAD, suggesting its potential as a diagnostic and prognostic biomarker in LUAD.
Humans ; Lung Neoplasms/genetics* ; Adenocarcinoma of Lung/genetics* ; China ; Prognosis ; Transcription Factors