Background/Aims: Natural killer (NK) cell function is generally considered dampened in chronic hepatitis B virus (HBV) infection; however, the NK cell pool exhibits phenotypic and functional heterogeneity, and the antibody--mediated effect of NK cells remains less characterized. This study evaluated the dynamic changes in antibody-mediated NK cell responses and the involvement of distinct NK subsets across disease stages and during antiviral treatment. Methods: A T-cell receptor-like antibody specific for the HBV core 18–27 peptide (cTCRL-Ab) was used to determine the antibody-mediated effect of NK cells, and an array of NK cell surface markers were analyzed in cross-sectional and longitudinal cohorts of patients with chronic HBV infection. Single-cell RNA sequencing (scRNA-seq) was performed to identify the heterogeneity of NK subsets. Results: The cTCRL-Ab enabled the detection of NK cell cytolytic activity and IFNγ production. Notably, cTCRL-Ab-mediated NK cell responses were compromised in chronically HBV-infected patients, particularly in those receiving pegylated interferon-α (Peg-IFNα), which was associated with the downregulation of CD16 expression. Correspondingly, Peg-IFNα inhibited cTCRL-Ab-mediated NK cell function by reducing CD16 expression in vitro. scRNA-seq revealed that CD16 downregulation occurred mainly within a dysfunctional CD16hi NK subset exhibiting exhaustion properties. In contrast, an activated CD16hiNK subpopulation (CX3CR1⁺KLRC2–CD16hi) with high cytotoxicity was enriched in patients who experienced favorable treatment responses. Furthermore, the intrahepatic CX3CR1+KLRC2–CD16hi subset tended to exhibit functional restoration in HBsAg-loss individuals. Conclusions Our data contribute to the understanding of antibody-mediated responses of NK cells in chronic HBV infection, and highlight a previously unappreciated functional CX3CR1+KLRC2–CD16hiNK subset as a potential therapeutic target.

[Objective]To investigate the association between screen content and the frequency of screen exposure at the age of one and a half years and hyperactive behavior in preschool,and to explore how the association is affected by the interaction between outdoor activities and screen behaviors,which could provide theoretical basis and feasible solutions for the prevention and intervention of behavioral problems in childhoood.[Methods]The survey was conducted from June 2022 to June 2023 in Huicheng District,Huizhou (China) stratified by whole cluster sampling methods. Parents and teachers of 5648 children in 61 kindergartens were sampled for questionnaire surveys. The Conners Teacher Rating Scale (TRS) was used to investigate hyperactive behavior. A self-administered questionnaire was used to investigate basic demographic information of children,screen content,frequency of screen exposure and outdoor activities at the age of one and a half years. Multivariate logistic regression was used to explore the association between video screen behavior and hyperactive behavior and its interaction with outdoor activities by controlling for covariates such as children's age,gender,and parental education.[Results]Result showed the overall prevalence of 3.2％ for hyperactive behavior,2.1％ for conduct problems,2.1％ for hyperactivity problems,1.3％ for inattention-passivity problems,and 0.9％ for hyperactivity index. After adjusting for confounding factors,multiple logistic regression analysis showed that screen exposure of "two to four times a week" at one and a half years old was associated with an increased detection rate of hyperactive behaviors in preschool children,with an estimated ORs (95％ CI) of 1.682 (1.141,2.480). Daily screen exposure was associated with increased detection rates of hyperactive behavior,conduct problems,hyperactivity issues,inattention-passivity problems,and hyperactivity index in pre-school age. The estimated ORs (95％ CI) were 2.136 (1.218,3.746),2.321 (1.185,4.546),2.300 (1.208,4.380),2.776 (1.267,6.085) and 3.640 (1.525,8.687),respectively. But the above associations were not found in children who were engaged in daily outdoor activities at the age of one and a half years (P value for interaction<0.001). No association was found between screen content and hyperactive behavior (P>0.05).[Conclusions]Frequency of screen exposure in early childhood is significantly associated with hyperactive behavior problems in preschool,and outdoor activities could weaken the correlation between high-frequency screen exposure and hyperactive behavior,suggesting that parents and schools should prioritize scientifically guiding children's video viewing behavior and outdoor activities,ensuring a well-arranged daily life,to lay a good foundation for the healthy development of children's behavior.

Objective To establish a project team to assess and manage the risk of out-of-town transport of extracorporeal membrane oxygenation(ECMO)patients using the healthcare failure mode and effects analysis(HFMEA),analyze the effectiveness of its application,and develop targeted improvement measures and processes.Methods Patients with ECMO who were treated in the department of intensive care unit(ICU)of Affiliated Hangzhou First People's Hospital,Zhejiang University School of Medicine March 2020 to September 2021 were selected as the study subjects.By comparing the differences in the accuracy of ECMO transport risk assessment,the rate of fixation against catheter slippage,the incidence of adverse transport events between the two groups before and after HFMEA,and the application effect of this model in ECMO transport adverse event risk management was evaluated.Results A total of 48 ECMO patients were enrolled,and 42 times of out-of-home transfers were performed,including 22 before HFMEA and 20 after HFMEA.Compared with pre-HFMEA,the accuracy of ECMO transport risk assessment[95.00%(19/20)vs.54.55%(12/22),P＜0.05]and the compliance rate of catheter slip fixation[100.00%(20/20)vs.68.18%(15/22),P＜0.05]in patients after HFMEA were significantly increased,the incidence of adverse events in transport was significantly lower[5.00%(1/20)vs.40.91%(9/22),P＜0.05].Conclusion The application of HFMEA in the risk management of ECMO transshipment adverse events is beneficial to standardize the prevention of ECMO transshipment accidents,effectively reduce the incidence of transshipment adverse events,and ensure patient safety.

Objective To investigate the application of venereal disease research laboratory(VDRL) test and toluidine red unheated serum test(TRUST) in syphilis laboratory diagnosis.Methods Serum,plasma and cerebrospinal fluid (CSF) in syphilis patients and healthy controls were measured by VDRL and TRUST.Results The VDRL detection results in serum and CSF sepcimens were generally higher than the TRUST detection results by 1-2 titers,while in plasma specimen,the VDRL detection results were generally lower than the TRUST detection results by 1-2 titers than TRUST when using plasma specimen.In addition,the VDRL detection in normal control serum and plasma specimens all appeared different degrees of false positive,but in the detection of normal control CSF,the results of TRUST and VDRL were consistent.Conclusion TRUST is more suitable for serum and plasma specimens,and CSF is suitable for the CSF specimen,but not suitable for serum and plasma detection.

Objective To realize the incidence of healthcare-associated infection(HAI)in patients after receiving extracorporeal membrane oxygenation(ECMO)procedure,and to evaluate the related factors for HAI.Methods Clinical data of patients receiving ECMO from January 2003 to December 2012 were collected and analyzed retro-spectively.Results Of 43 patients receiving ECMO,15 (34.88% )developed 24 times(55.81% )of HAI. The main HAI site was lower respiratory tract(n= 12,50.00% ),followed by blood stream(n= 6,25.00% ),skin and soft tis-sue(n= 5,20.83% ).A total of 28 isolates of pathogens were detected,gram-negative bacilli were 19(67.86% )iso-lates,gram-positive cocci 7(25.00% ),and fungi 2(7.14% );pathogens were mainly isolated from specimens of sputum(n= 12,42.86% ),blood (n= 9,32.14% )and wound secretion(n= 6,21 .43% ). The incidence of postopera-tive HAI in ECMO patients was related with patients’age,duration of ECMO,complication,mechanical ventila-tion,tracheal intubation or tracheotomy,and indwelling urinary catheter.Conclusion HAI in patients receiving EC-MO is high,hospital should take corresponding prevention and control measures targeting to the related risk factors of infection,so as to reduce the incidence of HAI after the ECMO.

Objective To assess the vitro susceptibility to 6 antimicrobial agents and genotypes of clinical isolates of Chlamydia trachomatis (Ct) from Guangzhou region. Methods Ct was isolated from clinical specimens by using McCoy cell culture and subjected to propagation. The minimum inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of 6 antimicrobial agents (clarithromycin, roxithromycin, azithromycin, doxycycline, tetracycline, ofloxacin) against Ct isolates were determined in McCoy cell culture. Nested PCR was performed to amplify the outer membrane protein 1 (omp1) VS1-2 gene followed by sequencing. Results Seventy-six Ct strains were isolated from 346 urogenital specimens, and 40 strains met the require ments for susceptibility testing after serial propagation. The MIC50/MIC90 of clarithromycin, azithromycin, roxi thromycin, doxycycline, tetracycline and ofloxacin were as follows: 0.008/0.032, 0.080/0.160, 0.125/0.500, 0.032/0.064, 0.250/0.500 and 0.500/1.000 mg/L. Seven genotypes were observed. The most prevalent geno types in decreasing order were E (14, 35%), J (10, 25%)and F (6, 15%). The MIC50 was consistent for azithromycin among the 7 genotypes, but varied by 1 - 4 folds for doxycycline, ofloxacin and roxithromycin. Conclusions Clarithromycin, doxycycline and azithromycin exhibit an excellent activity against Ct, and the activity of azithromycin is consistent among the 7 genotypes of Ct.

Objective To develop a PCR-mverse dot blot hybridization(RDB)assay to rapidly detect pathogenic mycoplasmas in genitourinary tract.Methods Universal primers were designed and applied to amplify the 16S rRNA gone of ureaplasma parvum(Up),ureaplasma urealyticum(Uu),Mycoplasma genitalium(Mg),Mycoplasma hominis(Mh)by using nestcd PCR.Specific nucleotide probes of Up,Uu,Mg and Mh Were constructed and immobilized onto nylon membranes.PCR products were denatured and hybridized、with specific oligonucleofide probes on nylon membrane.The sensitivity and specificity of the PCR-RDB assay were evaluated based.on the hybddizafion results.Also,PCR-RDB Was utilized to detect pathogenic mycoplasmas from 60 clinical samples.Results The four probes selectively hybridized with the PCR product of corresponding mycoplasmas,and no cross hybridization was observed.The detection limit of PCR-RDB Was one colony forming unit(CFU)of mycoplasma.Out of the 60 clinical samples、19were positive for mycoplasm,Mixed infections were found in three samples,including two coinfected with Up and Uu and one with Uu and Mg.Conclusion PCR-RDB is a rapid,specific and sensitive approach to the identification of pathogenic mycoplasmas in urogenital tract.

Objective To investigate abnormal glucose metablism of patients with stroke in hospital in Foshan in order to design more reasonable diagnostic and treatment plans.Methods Our study was conducted in 557 patients with stroke who were in hospital in Neurology Department of the First Hospital of Foshan from June 2007 to April 2008 (all were consistent with diagnostic criteria of stroke).Sex,age,history of diabetes mellitus and smoking,height,weight,blood pressure,waist circumference,random blood glucose,fasting blood glucose and oral glucose tolerance test of all were recorded when they were in hospital Results All 368 individuals (66.1%) had abnormal blood glucose,including 185 cases (33.2%) of diabetes,183 cases (32.9%) of impaired glucose tolerance and 189 cases (33.9%) of normal sugar tolerance.Furthermore,about 89.1% (189/368) cases with impaired glucose regulation and 14.1% (26/185) cases of diabetes were misdiagnosed if they did not undergo oral glucose tolerance test.Conclusions Most of patients with stroke in Foshan have impaired glucose regulation,who have been diagnosed by oral glucose tolerance test,in favour of us to better design more reasonable diagnostic and treatment plans.

The article aimed to explore the implementation of systemic integrated new clinical medical teaching mode and the functions of basic medicine specialty module teaching on innovative talents training.It showed that the implementation of systemic integrated new medical teaching mode was facilitated to build up a platform for the training of innovative talents.And the application of the integrated method to teaching was beneficial to develop the creative thinking of medical students.

Objective To develop a nested PCR for the detection of early syphilis and genotyping of Treponema pallidum (TP), and to investigate the distribution of genotypes of TP in Guangzhou. Methods Specimens were consecutively collected from genital ulcers of patients with suspected chancre during 2002-2004, and were detected by dark-field microscopy and nested PCR. The acidic repeat protein (arp) gene and the T. pallidum repeat (tpr) gene family were amplified with the positive specimens above. The number of repeats presented in the arp gene and the restriction fragment length polymorphism by Mse I in the tpr gene were analyzed by electrophoresis. The strains were genotyped according to Pillay's criteria. Results Out of 62 patients with suspected chancre, 33 cases (53.2%) were positive by dark-field microscopy and 54 cases (87.1%) by nested PCR. Of 47 TP-positive specimens genotyped by arp gene, 36 (76.6%) were type 14, while of 49 cases genotyped by tpr gene 39 (79.6%) were type d. By combining genotypes of arp and tpr genes, 7 genotypes were found, including 14d (31, 66.0%), 13d (5, 10.6%), 14b (4, 8.5%), 12b (3, 6.4%), 12d (2, 4.3%), 15d(l, 2.2%) and 14i (1, 2.2%). Conclusions Nested PCR shows a high sensitivity in early detection of TP. Genotype 14d seems the predominant type of TP in Guangzhou.