Objective To develop a method for detecting the monosaccharide composition of different serotypes of pneumococcal capsular polysaccharides(PnPs), in order to provide a reliable detection method for the quality control of pneumococcal PnPs vaccines and polysaccharide-protein conjugate vaccines.Methods According to the structural differences of different serotypes of PnPs, trifluoroacetic acid(TFA), hydrofluoric acid(HF) + TFA, and anhydrous hydrogen chloride/methanol(HCl/Meth) + TFA were used for hydrolysis to release the monosaccharide components, respectively. The high performance anionexchange chromatography with pulsed amperometric detection(HPAEC-PAD) without derivatization and 1-phenyl-3-methyl-5-pyrazolone(PMP) pre-column derivatization high-performance liquid chromatography(HPLC) were used for detection to compare the differences in the separation performance for acidic monosaccharides, neutral monosaccharides and alkaline monosaccharides between different detection methods. Different hydrilysis methods(TFA, HF + TFA, HCl/Meth + TFA, and HCl) were used to hydrolyze monosaccharides, and the stability of each monosaccharide component was evaluated by PMP pre-column derivatization HPLC.Results The three hydrolysis methods could effectively break the glycosidic bonds of different serotypes of PnPs and release various monosaccharide components. The PMP derivatization combined with a C18 column exhibited the best separation performance, enabling baseline separation of 10 neutral, acidic, and alkaline monosaccharides, suitable for the comprehensive analysis of all serotypes. In the HPAEC-PAD method, PA10 column had good separation performance for most monosaccharides, but some glycosamines were not well separated. MA1 column could specifically detect ribitol and glycerol components which cannot be derivatized, while could not elute uronic acids. HF and TFA treatments had little effect on the recovery of most monosaccharides. However, HCl/Meth treatment could easily lead to the degradation of hexosamine,while HCl treatment alone leaded to the degradation of neutral monosaccharides and uronicacids.Conclusion PMP pre-column derivatization HPLC has good separation performance and strong universality, and is a reliable method to detect the monosaccharide composition of different serotypes of PnPs. HPAEC-PAD method has unique advantages in detecting specific components such as ribitol. The combination of the two forms a complete method system for the quality control of pneumococcal polysaccharide conjugate vaccines.

Depression and circadian rhythms exhibit bidirectional interactions,suggesting a close association with the biological clock system.The biological clock in the suprachiasmatic nucleus of the hypothalamus is regulated by circadian genes.Recent clinical and basic research has revealed multifaceted associations between depression and circadian genes.For instance,significant phase abnormalities in BMAL1,PER2,and PER3 were detected in brain tissue from depressed patients,while plasma levels of CRY1,ARNTL,and PER1 proteins showed marked reduction,demonstrating good diagnostic value.Mice with CLOCK and BMAL1 knockouts exhibited depression-like behaviors.Single nucleotide polymorphisms(SNPs)in genes such as PER1 and PER3 directly influence depression susceptibility.Methylation levels of BMAL1,PER3,and CLOCK genes correlate closely with depressive symptoms.Antidepressant mechanisms like ketamine exert their effects by downregulating PER2 and other genes.This review summarizes the differential expression patterns of circadian clock genes in depression and associated therapeutic approaches,aiming to provide new theoretical foundations for precision diagnosis and personalized treatment strategies for depression.

Objective To conduct microbiological monitoring before the operation of the purified water treatment system in a newly-built endoscopy center,comprehensively analyze the causes for the standard-exceeding results of microbial detection of final rinse water,propose solutions,and provide reference for handling similar events in the future.Methods Microbial detection data of the final rinse water in the newly-built digestive endoscopy center of a tertiary first-class general hospital in Beijing from April to July 2024 were monitored.The potential causes for standard-exceeding results of microbial detection of final rinse water were analyzed from the perspectives of equip-ment maintenance and management of the purified water treatment system as well as the improvement of cleaning and disinfection methods for the purified water supply pipeline in the endoscopy center,targeted improvement mea-sures were proposed accordingly.Results The microbial monitoring result of final rinse water in the newly-built di-gestive endoscopy center built in April 2024 was 1 400 CFU/100 mL,with the main bacterial type being Cupriavi-duspauculus.After five rounds of improvement measures and rechecks,microbial monitoring result of the final rinse water in the newly-built endoscopy center was 0 CFU/100 mL,with a qualification rate of 100％.Analysis suggested that the main causes for the standard-exceeding results of microbial detection of final rinse water were due to the damage of the reverse osmosis membrane,lack of cleaning for the pure water storage tank before use,and non-standard cleaning and disinfection process for the pure water supply pipeline,after targeted improvement,the problem was solved.Conclusion Medical institutions should continuously conduct periodic monitoring on water used for endoscope,regularly perform cleaning and disinfection of the purified water treatment system,standardize cleaning and disinfection procedures,ensure medical quality and patient safety.

Objective:To explore the efficacy of non-invasive prenatal testing (NIPT) of fetal free DNA in maternal peripheral blood in fetuses with increased nuchal translucency (NT).Methods:A retrospective analysis was conducted on 1 184 singleton pregnant women that underwent chromosomal microarray analysis (CMA) at Nanjing Drum Tower Hospital, Nanjing University Medical School from June 2014 to December 2022 due to fetal increased NT (≥3.0 mm). These subjects were categorized based on whether the increased NT was accompanied by other high-risk factors into isolated increased NT without advanced maternal age (further subdivided into 3.0 mm≤NT<3.5 mm, 3.5 mm≤NT<4.0 mm, and NT≥4.0 mm subgroups), isolated increased NT with advanced maternal age, increased NT with nasal bone abnormalities, increased NT with other soft markers, and increased NT with structural abnormalities groups. Assuming the sensitivity and specificity of NIPT and expanded NIPT at this center were both 100%, genomic abnormalities outside the detection range of NIPT or expanded NIPT were termed as residual risk of NIPT or expanded NIPT. Chi-square test and Bonferroni correction were used to compare the residual risks of NIPT and expanded NIPT among the three subgroups of isolated increased NT without advanced maternal age group. Results:(1) In the group of isolated increased NT without advanced maternal age: For the 3.0 mm≤NT<3.5 mm subgroup (329 cases), 19 abnormalities were detected by CMA [12 cases of chromosome aneuploidy, seven cases of pathogenic copy number variation (pCNV)], with residual risks of NIPT and expanded NIPT both at 2.1% (7/329). For the 3.5 mm≤NT<4.0 mm subgroup (173 cases), 29 abnormalities were detected by CMA (17 cases of chromosome aneuploidy, nine cases of pCNV, three cases of chromosome unbalanced translocation), with residual risks of NIPT at 8.1% (14/173) and expanded NIPT at 7.5% (13/173). For the NT≥4.0 mm subgroup (270 cases), CMA detected abnormalities in 70 cases (50 cases of chromosome aneuploidy, 16 cases of pCNV, three cases of unbalanced translocations, and one case of sex chromosome abnormality combined with pCNV). The residual risk of NIPT was 12.2% (33/270), and the residual risk of expanded NIPT was 7.0% (19/270). The residual risks of NIPT and expanded NIPT in the 3.0 mm≤NT<3.5 mm subgroup were lower than those in the 3.5 mm≤NT<4.0 mm and NT≥4.0 mm subgroups (Bonferroni correction, all P<0.017). (2) In the group of 92 cases with isolated increased NT and advanced maternal age, CMA detected abnormalities in 36 cases (29 cases of chromosome aneuploidy, five cases of pCNV, one case of trisomy 21 combined with sex chromosome abnormality, and one case of trisomy 18 combined with sex chromosome abnormality). The residual risk of NIPT was 7.6% (7/92), and that of expanded NIPT was 5.4% (5/92). (3) In the group of 49 cases with increased NT combined with nasal bone abnormalities, CMA detected abnormalities in 24 cases (23 cases of chromosome aneuploidy and one case of pCNV). The residual risks of NIPT and expanded NIPT were both 2.0% (1/49). (4) In the group of 26 cases with increased NT combined with other soft markers, CMA detected abnormalities in nine cases (six cases of chromosome aneuploidy, one case of pCNV, and two cases of chromosome unbalanced translocations). The residual risks of NIPT and expanded NIPT were both 11.5% (3/26). (5) In the group of 245 cases with increased NT combined with structural abnormalities, CMA detected abnormalities in 121 cases (107 cases of chromosome aneuploidy, seven cases of pCNV, four cases of chromosome unbalanced translocations, one case of trisomy 21 combined with trisomy 20, and two cases of trisomy 18 combined with sex chromosome abnormalities). The residual risk of NIPT was 16.7% (41/245), and that of expanded NIPT was 4.1% (10/245). Conclusions:For isolated NT≥3.5 mm or NT≥3.0 mm combined with other high-risk factors, chorionic villus sampling in early pregnancy can be recommended, advancing the timing of prenatal diagnosis from the second trimester to the first trimester. For fetuses with isolated 3.0 mm≤NT<3.5 mm, the 2.1% residual risk of chromosomal abnormalities should be fully informed during counseling, even if the risk of NIPT is low.

Depression and circadian rhythms exhibit bidirectional interactions,suggesting a close association with the biological clock system.The biological clock in the suprachiasmatic nucleus of the hypothalamus is regulated by circadian genes.Recent clinical and basic research has revealed multifaceted associations between depression and circadian genes.For instance,significant phase abnormalities in BMAL1,PER2,and PER3 were detected in brain tissue from depressed patients,while plasma levels of CRY1,ARNTL,and PER1 proteins showed marked reduction,demonstrating good diagnostic value.Mice with CLOCK and BMAL1 knockouts exhibited depression-like behaviors.Single nucleotide polymorphisms(SNPs)in genes such as PER1 and PER3 directly influence depression susceptibility.Methylation levels of BMAL1,PER3,and CLOCK genes correlate closely with depressive symptoms.Antidepressant mechanisms like ketamine exert their effects by downregulating PER2 and other genes.This review summarizes the differential expression patterns of circadian clock genes in depression and associated therapeutic approaches,aiming to provide new theoretical foundations for precision diagnosis and personalized treatment strategies for depression.

Saliva is an important body fluid in the oral cavity containing lots of biomarkers, whose inherent micro-ecosystem holds significant value for early diagnosis and monitoring of oral diseases. Simultaneously, saliva has particular advantages, such as ease of sampling, painless and non-invasive collection, and suitability for repeated sampling, making it highly appropriate for surveillance and follow-up of diseases. In a series of studies conducted by the research group for preventive dentistry in Peking University School and Hospital of Stomatology, we compared different segments of saliva and those samples collected via different sampling methods using proteomic/peptidomic and microbiomic technologies to explore the stability of saliva samples. Besides, the significance of applying representative salivary biomarkers in early prevention and control of representative oral diseases (e.g. dental caries, periodontal diseases) and systemic conditions (e.g. type 2 diabetes mellitus, chronic kidney disease) was confirmed as well.
Humans ; Saliva/chemistry* ; Dental Caries/diagnosis* ; Biomarkers/analysis* ; Periodontal Diseases/diagnosis* ; Mouth Diseases/diagnosis* ; Proteomics/methods* ; Diabetes Mellitus, Type 2/diagnosis* ; Microbiota ; Renal Insufficiency, Chronic/prevention & control*

The spatio-temporal heterogeneity of breast cell subsets forms the fundamental biological basis for physiological development and pathological progression, including tumorigenesis; however, its complex regulatory mechanisms are not yet fully elucidated. With its high-resolution capabilities, single-cell RNA sequencing (scRNA-seq) technology offers a powerful tool for dissecting this cellular heterogeneity. This technology enables the construction of high-precision breast cell atlases, the accurate identification of distinct cell subsets, and the reconstruction of differentiation trajectories from stem/progenitor cells to functional epithelial cells. By resolving the transcriptional regulatory networks that govern cell fate determination, intercellular communication patterns, and dynamic microenvironmental interactions, scRNA-seq has unveiled the molecular foundations of breast development and provided new perspectives on the pathogenesis of related diseases such as breast cancer and macromastia. Furthermore, scRNA-seq demonstrates significant potential for discovering early molecular markers of disease, deciphering tumor heterogeneity, and elucidating mechanisms of therapeutic resistance. The continued application of scRNA-seq for dissecting breast cell heterogeneity, combined with its integration with multi-modal data such as spatial omics, promises to provide critical evidence and new insights for revealing the molecular mechanisms of breast development-related diseases and for formulating precision therapeutic strategies.
Humans ; Single-Cell Analysis/methods* ; Female ; Breast Neoplasms/pathology* ; Sequence Analysis, RNA/methods* ; Breast/cytology*

Objective:To systematically review the application of Artificial Intelligence (AI) in drug clinical trial management under the Good Clinical Practice (GCP) framework, identify current gaps and shortcomings, and to propose an optimization strategy.Methods:Main academic databases were searched for literature published in Chinese and English between September 2019 and September 2024. Relevant studies were included. Key findings were extracted and summarized.Results:AI applications had been extensively explored and reported in areas such as informed consenting and protection of human subject rights, trial design and implementation, subject screening and recruitment, data processing and management, and reporting. Current literature was predominantly international. Despite a foundation of applied research, issues existed such as lack of standardization, fragmented tools, and insufficient interconnectivity and interoperability.Conclusions:China has marked potential to adopt and integrate AI technologies in drug clinical trial management under the GCP framework. Future efforts should focus on establishing standards, covering end-to-end processes, promoting multi-tool collaboration and data interconnectivity, and building robust data security and ethical protection mechanisms in an AI environment. Seizing the opportunities can enhance drug clinical trial management.

Objective:To explore the efficacy of non-invasive prenatal testing (NIPT) of fetal free DNA in maternal peripheral blood in fetuses with increased nuchal translucency (NT).Methods:A retrospective analysis was conducted on 1 184 singleton pregnant women that underwent chromosomal microarray analysis (CMA) at Nanjing Drum Tower Hospital, Nanjing University Medical School from June 2014 to December 2022 due to fetal increased NT (≥3.0 mm). These subjects were categorized based on whether the increased NT was accompanied by other high-risk factors into isolated increased NT without advanced maternal age (further subdivided into 3.0 mm≤NT<3.5 mm, 3.5 mm≤NT<4.0 mm, and NT≥4.0 mm subgroups), isolated increased NT with advanced maternal age, increased NT with nasal bone abnormalities, increased NT with other soft markers, and increased NT with structural abnormalities groups. Assuming the sensitivity and specificity of NIPT and expanded NIPT at this center were both 100%, genomic abnormalities outside the detection range of NIPT or expanded NIPT were termed as residual risk of NIPT or expanded NIPT. Chi-square test and Bonferroni correction were used to compare the residual risks of NIPT and expanded NIPT among the three subgroups of isolated increased NT without advanced maternal age group. Results:(1) In the group of isolated increased NT without advanced maternal age: For the 3.0 mm≤NT<3.5 mm subgroup (329 cases), 19 abnormalities were detected by CMA [12 cases of chromosome aneuploidy, seven cases of pathogenic copy number variation (pCNV)], with residual risks of NIPT and expanded NIPT both at 2.1% (7/329). For the 3.5 mm≤NT<4.0 mm subgroup (173 cases), 29 abnormalities were detected by CMA (17 cases of chromosome aneuploidy, nine cases of pCNV, three cases of chromosome unbalanced translocation), with residual risks of NIPT at 8.1% (14/173) and expanded NIPT at 7.5% (13/173). For the NT≥4.0 mm subgroup (270 cases), CMA detected abnormalities in 70 cases (50 cases of chromosome aneuploidy, 16 cases of pCNV, three cases of unbalanced translocations, and one case of sex chromosome abnormality combined with pCNV). The residual risk of NIPT was 12.2% (33/270), and the residual risk of expanded NIPT was 7.0% (19/270). The residual risks of NIPT and expanded NIPT in the 3.0 mm≤NT<3.5 mm subgroup were lower than those in the 3.5 mm≤NT<4.0 mm and NT≥4.0 mm subgroups (Bonferroni correction, all P<0.017). (2) In the group of 92 cases with isolated increased NT and advanced maternal age, CMA detected abnormalities in 36 cases (29 cases of chromosome aneuploidy, five cases of pCNV, one case of trisomy 21 combined with sex chromosome abnormality, and one case of trisomy 18 combined with sex chromosome abnormality). The residual risk of NIPT was 7.6% (7/92), and that of expanded NIPT was 5.4% (5/92). (3) In the group of 49 cases with increased NT combined with nasal bone abnormalities, CMA detected abnormalities in 24 cases (23 cases of chromosome aneuploidy and one case of pCNV). The residual risks of NIPT and expanded NIPT were both 2.0% (1/49). (4) In the group of 26 cases with increased NT combined with other soft markers, CMA detected abnormalities in nine cases (six cases of chromosome aneuploidy, one case of pCNV, and two cases of chromosome unbalanced translocations). The residual risks of NIPT and expanded NIPT were both 11.5% (3/26). (5) In the group of 245 cases with increased NT combined with structural abnormalities, CMA detected abnormalities in 121 cases (107 cases of chromosome aneuploidy, seven cases of pCNV, four cases of chromosome unbalanced translocations, one case of trisomy 21 combined with trisomy 20, and two cases of trisomy 18 combined with sex chromosome abnormalities). The residual risk of NIPT was 16.7% (41/245), and that of expanded NIPT was 4.1% (10/245). Conclusions:For isolated NT≥3.5 mm or NT≥3.0 mm combined with other high-risk factors, chorionic villus sampling in early pregnancy can be recommended, advancing the timing of prenatal diagnosis from the second trimester to the first trimester. For fetuses with isolated 3.0 mm≤NT<3.5 mm, the 2.1% residual risk of chromosomal abnormalities should be fully informed during counseling, even if the risk of NIPT is low.