Glycoengineering was carried out in the mammalian cell line CHO for the production of pro-tein-based drugs.Firstly,the genome sequence of the Rosa26 locus of CHO cells was determined,the gRNA sequences were designed,and the landing pad was integrated into the Rosa26 locus of CHO cells by CRISPR/Cas9 technology.Three targeting vectors co-expressed by glycosyltransferases,which are β-1,4 galactosyltransferase(B4GALT1),α-2,6-sialyltransferase 1(ST6GAL1)and N-acetaminoglycosyl-transferase Ⅲ(GnT Ⅲ),were constructed by overlapping PCR and seamless ligation technology,and the three glycosyltransferase genes were integrated into the CHO Rosa26 locus by Cre enzyme-mediated cassette exchange technology.PCR confirmed that three glycosyltransferases had been successfully site-directed integrated into the Rosa26 site.The mRNA expression levels of the three glycosyltransferases were more than 50 000-fold by qRT-PCR,and the protein expression levels of the three glycosyltrans-ferases were more than 4-fold via western blotting(P＜0.001).A CHO-engineered cell line with three glycosyltransferases integrated into Rosa26 site was successfully constructed.

Objective:To report the nationwide surveillance results of pathogenic profiles and antimicrobial resistance patterns of Gram-positive bloodstream infections in China in 2023.Methods:The clinical isolates of Gram-posttive bacteria from blood cultures were collected in member hospitals of National Bloodstream Infection Bacterial Resistant Investigation Collaborative System（BRICS）during January to December 2023. Antimicrobial susceptibility testing was performed using the dilution method recommended by the Clinical and Laboratory Standards Institute（CLSI）. Statistical analyses were conducted using WHONET 5.6 and SPSS 25.0 software.Results:A total of 4 385 Gram-positive bacterial isolates were obtained from 60 participating center. The top five pathogens were Staphylococcus aureus（ n=1 544，35.2%），coagulase-negative Staphylococci（ n=1 441，32.9%）， Enterococcus faecium（ n=574，13.1%）， Enterococcus faecalis（ n=385，8.8%），and α-hemolytic Streptococci（ n=187，4.3%）. The prevalence of methicillin-resistant Staphylococcus aureus（MRSA）and methicillin-resistant coagulase-negative Staphylococci（MRCNS）was 26.2%（405/1 544）and 69.8%（1 006/1 441），respectively. Notably，all Staphylococci remained susceptible to glycopeptide or daptomycin. Staphylococcus aureus demonstrated excellent susceptibility（>97.0%）to cephalobiol，rifampicin，trimethoprim-sulfamethoxazole，linezolid，minocycline，tigecycline，and eravacycline. No Enterococcus exhibiting resistance to linezolid were detected. Glycopeptide resistance was uncommon but more frequent in Enterococcus faecium（resistance to vancomycin and teicoplanin：both 1.7%）compared to Enterococcus faecalis（both 0.3%）. The detection rates of MRSA and MRCNS exhibited significant regional variations across the country（ χ2=17.674 and 148.650，respectively，both P<0.001）. No vancomycin-resistant Enterococci were detected in central China. Institutional comparison demonstrated higher prevalence of MRSA（ χ2=14.111， P<0.001）and MRCNS（ χ2=4.828， P=0.028）in provincial hospitals than that in municipal hospitals. Socioeconomic analysis identified elevated detection rates of both MRSA（ χ2=18.986， P<0.001）and MRCNS（ χ2=4.477， P=0.034）in less developed regions（per capita GDP

Objective:To report the results of bacterial resistant investigation collaborative system（BRICS）on the distribution and antimicrobial resistance profile of clinical Gram-negative bacteria isolates from bloodstream infections in China in 2023，and provide reference for clinical tretment of bloodstream infections and prevention and control of bacterial resistance.Methods:The clinical isolates of Gram-negative bacteria from blood cultures in member hospitals of BRICS were collected during January 2023 to December 2023. Antibiotic susceptibility tests were conducted by agar dilution or broth dilution methods recommended by Clinical and Laboratory Standards Institute（CLSI）. WHONET 5.6 and SPSS 25.0 were used to analyze the data.Results:During the study period，11 492 strains of Gram-negative bacteria were collected from 60 hospitals，of which 10 098（87.9%）were Enterobacterales and 1 394（12.1%）were non-fermentative bacteria. The top 5 bacterial species were Escherichia coli（50.0%）， Klebsiella pneumoniae（26.1%）， Pseudomonas aeruginosa（5.1%）， Acinetobacter baumannii complex（5.0%）and Enterobacter cloacae complex（4.1%）. The ESBL-producing rates in Escherichia coli， Klebsiella pneumoniae and Proteus mirablilis were 46.8%（2 685/5 741），18.3%（549/2 999）and 44.0%（77/175），respectively. The prevalence of carbapenem-resistant Escherichia coli（CREC）and carbapenem-resistant Klebsiella pneumoniae（CRKP）were 1.3%（76/5 741）and 15.0%（450/2 999）；32.9%（25/76）and 78.0%（351/450）of CREC and CRKP were sensitive to ceftazidime/avibactam combination，respectively. 94.7%（72/76）and 90.2%（406/450）of CREC and CRKP were sensitive to aztreonam/avibactam combination. Furthermore，57.9%（44/76）and 79.1%（356/450）were sensitive to imipenem/relebactam combination. The prevalence of carbapenem-resistant Acinetobacter baumannii（CRAB）complex was 64.6%（370/573），while more than 80.0% of CRAB complex was sensitive to tigecycline，eravacycline and polymyxin B. The prevalence of carbapenem-resistant Pseudomonas aeruginosa（CRPA）was 17.0%（99/581）. There were differences in the composition ratio of Gram-negative bacteria in bloodstream infections and the prevalence of important Gram-negative bacteria resistance among different regions in China，with statistically significant differences in the prevalence of CREC，CRKP，CRPA and CRAB complex（ χ2=10.6，28.6，10.8 and 19.3， P<0.05）. The prevalence of ESBL-producing Escherichia coli， CREC，CRAB complex and CRKP were higher in provincial hospitals than those in municipal hospitals（ χ2=12.5，9.8，12.7 and 57.8，all P<0.01）. Conclusions:Gram-negative bacteria are the main pathogens causing bloodstream infections in China，and Escherichia coli is ranked in the top，while the trend of Klebsiella pneumoniae increases continuously with time. CRKP infection shows a slow upward trend，CREC infecton maintains a low prevalence level，and CRAB complex infection continues to exhibit a high prevalence rate. The composition and resistance patterns of pathogens causing bloodstream infections vary to some extent across different regions and levels of hospitals in China.

Objective:Previous studies have shown that insomnia is closely related to erectile dysfunction(ED).However,the causal relationship between them is still unclear.Mendelian randomization(MR)provides a new method for studying the relationship between the two,and the theory of heart-kidney interaction in TCM provides a new idea for exploring the causal relationship between them.Methods:Based on the statistical data collected by genome-wide association studies(GWAS),the causal relationship be-tween insomnia and ED was discussed by MR.Inverse variance weighted(IVW)is the main analysis method,and weighted median(WME),simple mode(SM),weighted mode(WM)and MR Egger method were the supplementary analysis to evaluate the causal effect.MR-Egger intercept test,Cochran Q test and leave-one-out method were used in sensitivity analysis to verify the reliability of MR results.Results:Thirty-nine SNPs significantly related to insomnia were finally included for MR analysis.The results of IVW method in MR analysis showed that insomnia had a significant causal relationship with the increased risk of ED(OR=3.111,95％CI=1.566-6.181,P=1.193 × 10-3).The results obtained by MR-Egger method,WME method,WM method and SM method were consistent with IVW method in the direction of effect.The sensitivity results suggested that the results of this study were robust.Conclusion:Our study reveals the causal relationship between insomnia and ED,which provides a new basis for future clinical practice and prevention and treatment of ED.

Objective:To analyze the current situation and hotspots of medication literacy research at home and abroad, and provide references for medication literacy research in China.Methods:The literature related to medication literacy in the Web of Science Core Collection Database, Scopus and China National Knowledge Infrastructure Database were retrieved (up to May 31, 2024). The CiteSpace software was used to analyze the number of published papers, countries, institutions, journals, authors and keywords, etc.Results:A total of 604 literature were included (361 in Chinese and 243 in English). The literature related to medication literacy were first seen in 2000, and the number grew slowly, which showed rapid growth after 2016, and reached a peak in 2023. The country with the largest number of published English literature was China (69 articles), followed by the United States (66 articles). The literature from the United States were cited 3 623 times, and those from China were cited 2 523 times. Central South University and the Third Xiangya Hospital of Central South University were tied for the first place in terms of the number of published articles as institutions (both 15 articles). The top 5 institutions in terms of the number of Chinese publications were Xiangya Third Hospital of Central South University, Central South University, Yanbian University, Affiliated Hospital of Yanbian University, and Tianjin Chest Hospital. The discipline with the largest number of published English literature was pharmacology/pharmacy (107 articles), followed by public environmental occupational health (88 articles) and general internal medicine (39 articles); the discipline with the largest number of Chinese published articles was clinical medicine (124 articles), followed by research on medical and health policies and regulations (56 articles), and medical education and marginal medical disciplines (34 articles). Keyword cluster analysis showed that the top 3 keywords in the English literature were medication errors, health education, and community pharmacy, while those in the Chinese literature were health literacy, self-management, and health education.Conclusions:Research on medication literacy has rapidly developed in recent years. China and the United States are the main countries for research related to medication literacy. Health education and medication errors are the mainstream of the research. Future research can focus on personalized assessment and intervention measures of medication literacy, so as to develop high-quality assessment tools for medication literacy.

In recent years, significant progress has been made in the treatment of chronic constipation, with high-quality studies emerging in areas such as fecal microbiota transplantation (FMT), sacral neuromodulation (SNM), acupuncture, and surgical techniques. The therapeutic approach is shifting from a traditional "medication and surgery" model toward an integrated strategy that includes dietary and defecation habit adjustment, FMT, SNM, acupuncture, pharmacotherapy, and surgery. Although FMT can partially improve stool frequency and consistency, its standardization and long-term efficacy require further validation. SNM demonstrates limited effectiveness in treating chronic constipation and is relatively cost-inefficient. Electroacupuncture remains controversial, though some studies support its value. Biofeedback therapy is recommended by multiple guidelines as the first-line treatment for dyssynergic defecation (DD), with portable home-based biofeedback systems showing considerable potential. For internal rectal prolapse (IRP) and rectocele (RC), various surgical options exist without a clear superiority, though laparoscopic ventral rectopexy (VMR) is increasingly favored due to its low recurrence rate and high patient satisfaction. In the surgical management of slow transit constipation (STC), total colectomy with ileorectal anastomosis remains the mainstream approach, while subtotal colectomy is gaining attention as an alternative.

Objective:To explore the characteristics of serum galactose-deficient IgA1 (Gd-IgA1) and Gd-IgA1/IgA ratio in normal children.Methods:The children from a kindergarten and 3 primary or secondary schools in Baoding (Hebei Province) from March to August 2023 were included in the cross-sectional study.According to their age, the children were divided into the 3-6-year-old group, >6-11-year-old group, and >11-16-year-old group.Besides, some adults were also included as the control group.The immunoturbidimetric assay was used to detect serum IgA levels, and the enzyme-linked immunosorbent assay (ELISA) was employed to detect Gd-IgA1 and Gd-IgA1/IgA levels.The above indicators were subject to normal distribution tests and analysis of variance (ANOVA)or rank sum test.Results:A total of 136 normal subjects were included for analysis.Among these subjects, there were 64 males and 72 females.There were 13 children in the 3-6-year-old group, 51 children in the >6-11-year-old group, and 56 children in the >11-16-year-old group.Besides, 16 normal adults (28-45 years old) from the physical examination center were also included as the control group.The serum IgA levels of the 3-6-year-old group, >6-11-year-old group, >11-16-year-old group, and adult group were (0.88±0.35) g/L, (1.38±0.65) g/L, (1.78±0.61) g/L, and (2.49±0.94) g/L, respectively.Except for no statistically significant difference between the 3-6-year-old group and the >6-11-year-old group, as well as between the >11-16-year-old group and the adult group, all other age groups showed statistically significant differences in serum IgA levels (all P<0.05).The serum Gd-IgA1 levels of the 3-6-year-old group, >6-11-year-old group, >11-16-year-old group, and adult group were 7.77(5.77, 16.51) μg/L, 7.12(5.29, 9.82) μg/L, 8.96(6.21, 14.30) μg/L, and 9.39(6.63, 40.00) μg/L, respectively, with statistically significant differences ( H=9.22, P=0.027).There was no significant difference in the Gd-IgA1/IgA ratio among different groups ( P=0.130).The correlation analysis suggested that Gd-IgA1/IgA1 did not correlate with age ( r=0.134, P=1.200).The Gd-IgA1 level was positively correlated with the IgA level ( r=0.204, P=0.017).There was no significant difference in serum IgA, Gd-IgA1, and Gd-IgA1/IgA levels between different gender groups. Conclusions:There are differences in Gd-IgA1 levels among different age groups among these children, but Gd-IgA1/IgA is not correlated with age.As a diagnostic biomarker, Gd-IgA1 should be emphasized from the different levels among different age groups, while Gd-IgA1/IgA seems to be more convenient for clinical research and application.

Oral squamous cell carcinoma(OSCC)is a malignant lesion originating from the oral mucosal squamous epithelium,account-ing for over 80％of oral and maxillofacial malignancies.Key etiological factors include tobacco,alcohol abuse,and betel quid chewing.In China,its incidence has shown an overall upward trend,posing a significant threat to public health.OSCC exhibits high local invasive-ness,making early diagnosis critical for improving prognosis.Its clinical management requires close multidisciplinary collaboration among oral and maxillofacial surgery,head and neck surgery,radiation oncology,medical oncology,reconstructive surgery,radiology,patholo-gy,and nutritional support teams.Given the increasing disease burden of OSCC and rapid development of multidisciplinary collaborative models,an expert panel has formulated this integrated management consensus based on evidence-based medicine and extensive deliber-ation.Centered on the'Prevention-Screening-Diagnosis-Treatment-Rehabilitation'framework,the consensus provides comprehensive guidance for the entire disease course of OSCC patients,aiming to standardize clinical practice.

Oropharyngeal squamous cell carcinoma(OPSCC)is a malignant tumor originating from the squamous epithelium of the oro-pharyngeal mucosa,accounting for more than 90％of oropharyngeal malignancies.In recent years,human papillomavirus(HPV)infec-tion has become one of the primary etiological factors of oropharyngeal squamous carcinoma.The incidence of HPV-associated oropharyn-geal squamous carcinoma has been rising annually,with a noticeable trend toward younger populations,posing a significant threat to hu-man health.Due to the distinct biological behavior and clinical characteristics of HPV-associated oropharyngeal squamous carcinoma com-pared to its non-HPV-related counterpart,the diagnostic and treatment strategies for oropharyngeal squamous carcinoma have undergone substantial changes.Prevention and screening for oropharyngeal squamous carcinoma are of critical importance.The diagnostic and treat-ment process involves multi-disciplinary collaboration,including oral and maxillofacial surgery,otolaryngology,head and neck surgery,oncology,radiology and pathology.Based on evidence from clinical practice,a comprehensive,integrated diagnostic and therapeutic ap-proach has been established,centered around the concept of"prevention,screening,diagnosis,treatment,and rehabilitation",covering the entire patient lifecycle and providing a valuable reference for clinical practice.

Glycoengineering was carried out in the mammalian cell line CHO for the production of pro-tein-based drugs.Firstly,the genome sequence of the Rosa26 locus of CHO cells was determined,the gRNA sequences were designed,and the landing pad was integrated into the Rosa26 locus of CHO cells by CRISPR/Cas9 technology.Three targeting vectors co-expressed by glycosyltransferases,which are β-1,4 galactosyltransferase(B4GALT1),α-2,6-sialyltransferase 1(ST6GAL1)and N-acetaminoglycosyl-transferase Ⅲ(GnT Ⅲ),were constructed by overlapping PCR and seamless ligation technology,and the three glycosyltransferase genes were integrated into the CHO Rosa26 locus by Cre enzyme-mediated cassette exchange technology.PCR confirmed that three glycosyltransferases had been successfully site-directed integrated into the Rosa26 site.The mRNA expression levels of the three glycosyltransferases were more than 50 000-fold by qRT-PCR,and the protein expression levels of the three glycosyltrans-ferases were more than 4-fold via western blotting(P＜0.001).A CHO-engineered cell line with three glycosyltransferases integrated into Rosa26 site was successfully constructed.