ObjectiveTo investigate the change in the activity of hepatitis B virus （HBV）-specific CD8⁺ T cells after pegylated interferon-α-2b （PEG-IFN-α-2b） therapy in HBeAg-negative patients with chronic HBV infection. MethodsA total of 53 HBeAg-negative patients with chronic HBV infection who attended The First Affiliated Hospital of Xinxiang Medical University and Tangdu Hospital of Air Force Mdical University from April 2020 to June 2022 were enrolled and treated with PEG-IFN-α-2b （180 μg/week， subcutaneous injection） antiviral therapy. The study endpoint was HBsAg clearance （course of treatment<48 weeks） or 48 weeks （course of treatment≥48 weeks）. Peripheral blood mononuclear cells were isolated at baseline and study endpoint， and peripheral blood T cell counts were measured. Enzyme-linked immunospot assay was used to measure the frequency of HBV-specific CD8⁺ T cells secreting perforin， granzyme B， and interferon-γ. A total of 17 HLA-A^*02-restricted patients were selected， and CD8⁺ T cells were purified to establish direct- and indirect-contact co-culture systems for HBV-specific CD8⁺ T cells and HepG2.2.15 cells. The level of lactate dehydrogenase in supernatant was measured to calculate the mortality rate of HepG2.2.15 cells， and the levels of HBV DNA， cytotoxic molecules， and cytokines in supernatant were also measured. Flow cytometry was used to measure the expression of apoptosis ligands， and the cytotoxicity of HBV-specific CD8⁺ T cells was evaluated. The independent samples t-test or the paired t-test was used for comparison of normally distributed continuous data between two groups， and the Mann-Whitney U test or the Wilcoxon rank-sum test was used for comparison of non-normally distributed continuous data between two groups. ResultsThe HBsAg clearance rate at study endpoint was 30.19% （16/53）. There were no significant differences in peripheral blood T cell counts （CD3⁺， CD4⁺， and CD8⁺ T cells） between baseline and study endpoint （P>0.05）. At study endpoint， there was a significant increase in the frequency of HBV-specific CD8⁺ T cells secreting perforin， granzyme B， and interferon-γ （U=177.50， t=11.90， U=186.50， all P<0.001）， and the patients with HBsAg clearance had a significantly higher frequency of such HBV-specific CD8⁺ T cells than those without HBsAg clearance （U=120.50， t=2.73， U=121.50， all P<0.01）. In the direct- and indirect-contact co-culture systems at study endpoint， HBV-specific CD8⁺ T cells induced a significant reduction in HBV DNA in the supernatant of HepG2.2.15 cells （all P<0.001） and significant increases in the secretion of interferon-γ and tumor necrosis factor-α （all P<0.05）； in the direct-contact co-culture system， HBV-specific CD8⁺ T cells induced significant increases in the mortality rate of HepG2.2.15 cells （13.62%±3.27% vs 11.39%±2.40%， t=2.27， P=0.030） and the secretion of perforin and granzyme B （t=72.50， U=52.50， both P<0.05）. In the direct- and indirect-contact co-culture systems， compared with HBV-specific CD8⁺ T cells from the patients without HBsAg clearance， the HBV-specific CD8⁺ T cells from patients with HBsAg clearance had a significantly greater reduction in HBV DNA （P<0.05） and significant increases in the secretion of interferon-γ and tumor necrosis factor-α （P<0.05）. ConclusionPEG-IFN-‍α‍-2b therapy can help to achieve a relatively high HBsAg clearance rate in HBeAg-negative patients with chronic HBV infection， and the activity of HBV-specific CD8⁺ T cells is significantly enhanced， which is closely associated with HBsAg clearance.

ObjectiveWith the epidermal growth factor receptor（EGFR）/Signal Transducers and Activators of Transcription（STAT3）/Hexokinase 2（HK2） signaling pathway in atherosclerosis （AS） and ovarian cancer （OC） as the entry point， this paper discusses the molecular mechanism of Gypenoside L （Gyp-L） treating AS and OC with different diseases， provides a new perspective and theoretical basis for TCM treating AS and OC with EGFR-STAT3-HK2 pathway， and enriches the scientific connotation of the theory of "cytoskeleton in the heart". MethodsCCK-8 was used to detect the proliferation of HUVEC and OVCAR-3 cells， in order to determine the intervention concentration for subsequent experiments. The colorimetric method was used to detect the NO content in HUVEC and the contents of pyruvate and LDH in two cell lines. Cell cloning experiments and scratch experiments reflect the proliferation and migration ability of OVCAR-3 cells. Western blot was used to detect the expression levels of relevant proteins. Furthermore， two cell models overexpressing EGFR were constructed and co treated with Gyp-L. HUVEC cells were divided into control， ox-LDL， OE-NC， OE-EGFR， OE-NC+Gyp-L， and OE-EGFR+Gyp-L group. OVCAR-3 cells were divided into control， OE-NC， OE-EGFR ， OE-NC+Gyp-L， and OE-EGFR+Gyp-L group. The colorimetric method was used to detect the NO content in HUVEC and the contents of pyruvate and LDH in two cell lines. Western blot was used to detect the expression levels of EGFR-STAT3-HK2 pathway related proteins. Cell cloning experiments and scratch experiments reflect the proliferation and migration ability of OVCAR-3 cells. ResultsGyp-L can significantly reduce the NO content of HUVEC and the pyruvate and LDH content of two cell lines （P<0.05）； Inhibit the proliferation and migration ability of OVCAR-3 cells； Reduce the expression levels of EGFR/STAT3/HK2 pathway related proteins in HUVEC and OVCAR-3 cell lines （P<0.05）， and inhibit the glycolysis pathway. ConclusionGyp-L can inhibit glycolysis in HUVEC and OVCAR-3 cells through the EGFR/STAT3/HK2 pathway，thereby suppressing the occurrence and development of AS and OC.

ObjectiveWith the epidermal growth factor receptor（EGFR）/Signal Transducers and Activators of Transcription（STAT3）/Hexokinase 2（HK2） signaling pathway in atherosclerosis （AS） and ovarian cancer （OC） as the entry point， this paper discusses the molecular mechanism of Gypenoside L （Gyp-L） treating AS and OC with different diseases， provides a new perspective and theoretical basis for TCM treating AS and OC with EGFR-STAT3-HK2 pathway， and enriches the scientific connotation of the theory of "cytoskeleton in the heart". MethodsCCK-8 was used to detect the proliferation of HUVEC and OVCAR-3 cells， in order to determine the intervention concentration for subsequent experiments. The colorimetric method was used to detect the NO content in HUVEC and the contents of pyruvate and LDH in two cell lines. Cell cloning experiments and scratch experiments reflect the proliferation and migration ability of OVCAR-3 cells. Western blot was used to detect the expression levels of relevant proteins. Furthermore， two cell models overexpressing EGFR were constructed and co treated with Gyp-L. HUVEC cells were divided into control， ox-LDL， OE-NC， OE-EGFR， OE-NC+Gyp-L， and OE-EGFR+Gyp-L group. OVCAR-3 cells were divided into control， OE-NC， OE-EGFR ， OE-NC+Gyp-L， and OE-EGFR+Gyp-L group. The colorimetric method was used to detect the NO content in HUVEC and the contents of pyruvate and LDH in two cell lines. Western blot was used to detect the expression levels of EGFR-STAT3-HK2 pathway related proteins. Cell cloning experiments and scratch experiments reflect the proliferation and migration ability of OVCAR-3 cells. ResultsGyp-L can significantly reduce the NO content of HUVEC and the pyruvate and LDH content of two cell lines （P<0.05）； Inhibit the proliferation and migration ability of OVCAR-3 cells； Reduce the expression levels of EGFR/STAT3/HK2 pathway related proteins in HUVEC and OVCAR-3 cell lines （P<0.05）， and inhibit the glycolysis pathway. ConclusionGyp-L can inhibit glycolysis in HUVEC and OVCAR-3 cells through the EGFR/STAT3/HK2 pathway，thereby suppressing the occurrence and development of AS and OC.

Objective: To investigate the intentions of mothers of ageappropriate girls in Qingdao to vaccinate their daughters against human papillomavirus (HPV), so as to provide theoretical guidance for targeted health education in the future. Methods: A multistage random sampling method was adopted to conduct a crosssectional study among 2 244 mothers of girls aged 12-14 years in Qingdao from March to December 2023. The Mann-Whitney U test was used for group comparisons, and Logistic regression was performed to analyze the factors that influenced maternal intention to vaccinate their ageappropriate daughters against HPV. Results: Among the surveyed mothers, 89.22% (n=2 002) intended to vaccinate their daughters against HPV, and 68.58% (n=1 539) had fully vaccinated or had plans to complete it for themselves. The knowledge score of mothers intended to vaccinate their daughters was 10 (8, 11). The multivariate Logistic regression analysis showed that mothers aged >45 years (OR=0.19), those with an annual family income of 60 000-<150 000 yuan (OR=0.65), 150 000-<300 000 yuan (OR=0.58), 300 000-500 000 yuan (OR=0.22), and those with higher knowledge scores (OR=0.90) were more likely to vaccinate their daughters (P<0.05). Mothers with a junior college or undergraduate degree (OR=1.66), those who never or occasionally screened for HPV (OR=1.58), those who were intended to be vaccinated, not planning to complete the fullcourse vaccination, or overaged and unvaccinated (OR=7.13), those who were not concerned about their daughters HPV infection (OR=2.54), and those whose daughters were not in adolescence (OR=1.93) were less intended to vaccinate their daughters (P<0.05). The primary reasons for vaccine hesitancy were vaccine safety concerns (65.06%), followed by the belief of mothers that "the children is to young, and can be vaccinated when they are older" (13.25%). Conclusions Mothers of eligible girls in Qingdao have relatively higher intentions to vaccinate their daughters against HPV, and willingness is influenced by factors such as the mothers vaccination status, knowledge level, and daughters development stage. It is recommended to strengthen targeted health education, improve the cognitive level and acceptance of mother, and increase the vaccination rate of HPV vaccines.

Objective: To investigate the epidemiological and spatio-temporal characteristics of influenza in Jiaxing City, Zhejiang Province, so as to provide insights into perfecting the prevention and control strategies of influenza. Methods: Data of influenza in Jiaxing City from 2019 to 2023 were collected from the Chinese Disease Prevention and Control Information System. Population data of the same period were collected from the Zhejiang Health Information Network Reporting System. The epidemiological characteristics of influenza were analyzed using descriptive analysis. Vector map information was collected from the Open Street Map, and the spatio-temporal clustering characteristics of influenza were analyzed using spatial autocorrelation and spatio-temporal scanning. Results: A total of 181 501 cases of influenza were reported in Jiaxing City from 2019 to 2023, with an average annual reported incidence of 653.93/105. The majority of cases were aged 5 to <15 years (59 785 cases, 32.94%). The majority of the occupations were students (78 239 cases, 43.11%) and pre-school children (33 715 cases, 18.58%). The county (city, district) with the highest reported incidence was Haining City (1 451.70/105), and the town (street) with the highest reported incidence was Chang'an Town (1 932.78/105). Spatial autocorrelation analysis showed that the incidence of influenza in Jiaxing City from 2019 to 2023 had positive spatial correlations (all Moran's I>0, all P<0.05), with a high-high clustering in the southern region. Spatio-temporal scanning analysis showed that there was a spatio-temporal clustering of influenza in Jiaxing City from 2019 to 2023, with the southern region being the primary-type clustering area and the period between November and January of the following year being the clustering time. Conclusion There was a significant spatio-temporal clustering of influenza in Jiaxing City from 2019 to 2023, with winter being the peak season and the southern region being the primary area.

Objective : To analyze the epidemiological characteristics and spatial clustering of brucellosis in Shanxi Province from 2019 to 2023, so as to provide a reference for formulating prevention and control measures of brucellosis. Methods: The case data of brucellosis in Shanxi Province from 2019 to 2023 were collected through the Infectious Disease Surveillance System of the Chinese Disease Prevention and Control Information System. The seasonal distribution, population distribution, and region distribution of brucellosis cases were described. Spatial autocorrelation analysis was applied to explore the spatial clustering characteristics of brucellosis. Results: A total of 21 241 human brucellosis cases were reported in Shanxi Province from 2019 to 2023, with an average annual reported incidence of 11.87/100 000, showing an upward trend (P<0.05). The peak incidence period was from March to August, with 14 163 cases reported cumulatively, accounting for 66.68% of the total. There were 16 336 male cases and 4 905 female cases, with a male-to-female ratio of 3.33:1. The high-incidence age group was 40-<70 years, with 15 675 cases accounting for 73.80%. The majority of patients were farmers, with 17 926 cases accounting for 84.39%. Spatial autocorrelation analysis showed that there was spatial clustering in the incidence of brucellosis from 2019 to 2023 (all Moran's I>0, P<0.05). The high-high clustering areas were mainly Datong City, and Shuozhou City in northern Shanxi, and Linfen City in the southern Shanxi. The low-low clustering areas were mainly Taiyuan City and Yangquan City in central Shanxi, and Changzhi City and Jincheng City in southeastern Shanxi. Conclusions From 2019 to 2023, the reported incidence of brucellosis in Shanxi Province showed an upward trend. The incidence peaked from March to August, and males, middle-aged and elderly people and farmers were the high-risk groups. There was spatial clustering and the high-high clustering areas gradually expanded from northern Shanxi to southern Shanxi.

Objective To monitor the susceptibility of clinical isolates to antimicrobial agents in tertiary hospitals in major regions of China in 2022.Methods Clinical isolates from 58 hospitals in China were tested for antimicrobial susceptibility using a unified protocol based on disc diffusion method or automated testing systems.Results were interpreted using the 2022 Clinical &Laboratory Standards Institute(CLSI)breakpoints.Results A total of 318 013 clinical isolates were collected from January 1,2022 to December 31,2022,of which 29.5％were gram-positive and 70.5％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)was 28.3％,76.7％and 77.9％,respectively.Overall,94.0％of MRSA strains were susceptible to trimethoprim-sulfamethoxazole and 90.8％of MRSE strains were susceptible to rifampicin.No vancomycin-resistant strains were found.Enterococcus faecalis showed 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 94.2％in the isolates from children and 95.7％in the isolates from adults.The resistance rate to carbapenems was lower than 13.1％in most Enterobacterales species except for Klebsiella,21.7％-23.1％of which were resistant to carbapenems.Most Enterobacterales isolates were highly susceptible to tigecycline,colistin and polymyxin B,with resistance rates ranging from 0.1％to 13.3％.The prevalence of meropenem-resistant strains decreased from 23.5％in 2019 to 18.0％in 2022 in Pseudomonas aeruginosa,and decreased from 79.0％in 2019 to 72.5％in 2022 in Acinetobacter baumannii.Conclusions The resistance of clinical isolates to the commonly used antimicrobial agents is still increasing in tertiary hospitals.However,the prevalence of important carbapenem-resistant organisms such as carbapenem-resistant K.pneumoniae,P.aeruginosa,and A.baumannii showed a downward trend in recent years.This finding suggests that the strategy of combining antimicrobial resistance surveillance with multidisciplinary concerted action works well in curbing the spread of resistant bacteria.

Objective:To investigate the efficacy of Peritoneal interposition flap (PIF) technique in preventing postoperative pelvic lymphocele formation during laparoscopic radical prostatectomy with extended pelvic lymph node dissection (LRP+ ePLND).Methods:A retrospective analysis was conducted on clinical data of 113 patients with locally high-risk or locally advanced prostate cancer who underwent LRP+ ePLND at Shanghai East Hospital, from January 2020 to November 2023. Among them, 27 patients received PIF technique and 86 received traditional LRP+ ePLND. ePLND was carried out as the clearance of external iliac vessels, medial side of the internal iliac artery, and pararectal lymph nodes. The PIF technique was the suturing the peritoneal flap after freeing the bladder to the lateral side of the bladder, pulling the peritoneal edge that follows the bladder's free edge posteriorly to the pubis, curling it onto the lateral surface of the bladder. This could expose the lymph node clearance bed, establishing a pathway from the lymph node clearance bed to the abdominal cavity space, allowing exuded lymphatic fluid to flow into the abdominal cavity for absorption by the peritoneum. There were no statistically significant differences in age [(68.37±6.92)years vs.(70.47±5.72)years], body mass index [(25.47±2.49)kg/m 2vs.(24.46±2.80)kg/m 2], and preoperative PSA [(23.28±13.94)ng/ml vs.(24.81±13.99)ng/ml] between the PIF group and the control group ( P>0.05). Biopsy Gleason score in PIF group: 6 in 2 cases, 7 in 9 cases, 8 in 9 cases, 9-10 in 2 cases. Biopsy Gleason score in control group: 6 in 4 cases, 7 in 35 cases, 8 in 27 cases, 9-10 in 20 cases. Clinic stage in PIF group: T 2 in 18 cases, T 3 in 6 cases, T 4 in 3 cases. Clinic stage in control group: T 2 in 51cases, T 3 in 27 cases, T 4 in 8 cases. The preoperative Gleason scores and TNM staging comparisons between the PIF group and the control group showed no statistically significant differences ( P>0.05). Surgical duration, intraoperative blood loss, lymph node positivity rate, incidence of postoperative lymphocele, and recovery of urinary control were compared between the two groups. Results:All surgeries were completed successfully without intraoperative complications in both groups. There were no statistically significant differences between the PIF group and the control group in terms of surgical duration [(202.96±24.15)min vs.(201.1±29.85)min], intraoperative blood loss [(85.56±32.27)ml vs.(90.7±49.25)ml], and lymph node positivity rate [(4 in PIF group, 14.8%)vs.(25 in control group, 29.1%)]( P>0.05). Urinary catheters were retained for 10-14 days postoperatively. Following catheter removal, there were no statistically significant differences in urinary control rates at 1 month [51.85%(14/27)vs. 48.83%(42/86)]and 2 months[74.07%(20/27) vs. 72.09%(62/86)] between the PIF group and the control group ( P>0.05). At the 2 to 6-month follow-up CT scan, none of the 27 patients in the PIF group developed pelvic lymphocele, whereas 9 patients in the control group did (6 cases bilateral, 3 cases unilateral), showing a statistically significant difference between the two groups ( P=0.002). Postoperatively, 3 patients in the control group experienced symptoms, with 1 case of lymphocele infection causing fever 1 month after surgery. Lymphocysts were found in 2 patients with ipsilateral lower extremity swelling 2 weeks after surgery. Conclusions:The application of PIF technique during laparoscopic radical prostatectomy with extended pelvic lymph node dissection via the abdominal approach could be safe and feasible. It may prevent postoperative pelvic lymphocele formation.

Purpose/Significance To investigate the effectiveness of experiential teaching in perinatal healthcare management contin-uing education.Method/Process The perinatal healthcare team of Peking Union Medical College Hospital holds a four-day experiential learning program for 30 maternity-related medical staffs from China.Through the self-administered experiential teaching questionnaire(ETQ)and satisfaction questionnaire,the researchers analyze the participants'post-teaching skill acquisition,clinical application abil-ity and educational needs,as well as their satisfaction with course content,course duration,teaching design,classroom atmosphere,and teaching format,which are quantified with Likert scale.Result/Conclusion The experiential teaching mode shows good teaching effects in perinatal healthcare management continuing education,and has higher application value.In the future,the researchers should combine it with theoretical teaching and carry out more comprehensive and structured periodic learning classes throughout the country,so as to im-prove both the theoretical foundation and practical ability of healthcare workers for better healthcare services for pregnant women.

Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.