Sennoside A (SA), a typical prodrug, exerts its laxative effect only after its transformation into rheinanthrone catalyzed by gut microbial hydrolases and reductases. Hydrolases have been identified, but reductases remain unknown. By linking a photoreactive group to the SA scaffold, we synthesized a photoaffinity probe to covalently label SA reductases and identified SA reductases using activity-based protein profiling (ABPP). From lysates of an active strain, Bifidobacterium pseudocatenulatum (B. pseudocatenulatum), 397 proteins were enriched and subsequently identified using mass spectrometry (MS). Among these proteins, chromate reductase/nicotinamide adenine dinucleotide (NADH) phosphate (NADPH)-dependent flavin mononucleotide (FMN) reductase/oxygen-insensitive NADPH nitroreductase (nfrA) was identified as a potent SA reductase through further bioinformatic analysis and The Universal Protein Resource (UniProt) database screening. We also determined that recombinant nfrA could reduce SA. Our study contributes to further illuminating mechanisms of SA transformation to rheinanthrone and simultaneously offers an effective method to identify gut bacterial reductases.

Objective: To understand the epidemiological and etiological characteristics of food poisoning event occurred in a school in Hangzhou, Zhejiang, so as to provide reference for the scientific management of related emergencies. Methods: By determining the nature of the event through epidemiological investigation, a case control study was carried out to spot suspicious food in May 2024. The hygienic investigation was conducted to find out possible pollution links and factors, patients and canteen practitioners anal swab, canteen retention samples, catering link daub and other specimens were collected ,for rapid pathogen screening. And the suspected pathogen Clostridium perfringens (CP) were isolated and identified according to the screening results, and toxin gene detection and whole genome sequencing and cluster analysis of CP isolated strains were carried out. Results: The incident resulted in 45 people experiencing gastrointestinal symptoms, such as diarrhea and abdominal pain. The suspicious food was tomato scrambled eggs and corn ribs provided by the student canteen for lunch on May 29. A hygiene investigation found that there was a risk of contamination in the food processing, preparation and storage. A total of 46 anal swabs and 10 canteen retention samples were positive for CP 16 S, 59 strains of CP were isolated from 27 samples, 10 cases and 1 practitioner isolate were positive for CPE ( cpe ) (F mode), and their whole genome evolution analysis was conducted based on the same source. Conclusions The food poisoning event is caused by CP infection carrying CPE ( cpe ) (F mode), and the possible sources of outbreak are the carriers of the CP by employees. It is recommended that cafeteria staff strengthen training on common foodborne diseases and conduct regular monitoring of pathogens.

Interfering hepatitis B virus (HBV) capsid assembly holds promise as a therapeutic approach for chronic hepatitis B (CHB). Novel anti-HBV agents are urgently needed to overcome drug resistance challenges, with targeted protein degradation (TPD) emerging as a hopeful strategy. Herein, we report the first degradation of HBV core protein (HBC), a multifunctional structural protein, using small-molecule degraders developed by hydrophobic tagging (HyT) technology. Structure-activity relationship (SAR) analysis identified compound HyT-S7, featuring an adamantyl group, exhibiting potent inhibitory activity (EC₅₀ = 0.46 μmol/L, HepAD38 cells) and degradation ability (DC₅₀ = 3.02 ± 0.54 μmol/L) in a dose- and time-dependent manner. Mechanistic studies demonstrated that the autophagy-lysosome pathway was a potential driver of HyT-S7-induced HBC degradation. Remarkably, HyT-S7 effectively degraded 11 drug-resistant mutants, including highly resistant strains P25G and T33N, to Phase III drug GLS4. Furthermore, cellular thermal shift assay, surface plasmon resonance assay, and molecular dynamics simulations revealed the precise mode of HyT-S7 binding to HBC with the adamantyl group potentially mimicking protein misfolding to facilitate HBC degradation. This first proof-of-concept study highlights the potential of HyT-mediated TPD in HBC as a promising avenue for discovering novel HBV and other antiviral agents with favorable drug resistance profiles.

OBJECTIVE To evaluate the clinical value of dose-escalated postoperative radiotherapy(PORT)in improving local control and survival outcomes for head and neck adenoid cystic carcinoma(ACC)patients.METHODS This retrospective study analyzed 336 ACC patients treated with surgery plus PORT at Beijing Tongren Hospital from January 2015 to January 2021.Cohort stratification compared high-dose(>60 Gy,n=146)and conventional-dose(≤60 Gy,n=190)regimens.Survival analysis employed Kaplan-Meier estimates with log-rank testing,complemented by multivariate Cox regression for risk adjustment.RESULTS The cohort demonstrated 39.29%(132/336)cumulative local failure rate.The overall survival rates at 1,3,and 5 years after surgery were 98.81%,94.05%,and 90.48%,respectively.Dose-response relationships revealed:1.Significantly reduced local recurrence with high-dose PORT(28.08%vs.47.89%,P<0.001),corresponding to 41.37%lower recurrence risk(a HR=0.59,95%CI=0.38-0.91;P=0.041);2.Superior progression-free survival in the high-dose group(3-year:86.99%vs.76.32%;5-year:82.19%vs.66.32%,all P<0.05);3.Comparable overall survival between groups(median OS:200 vs.160 months,P=0.292).CONCLUSION Dose escalation beyond 60 Gy significantly enhances locoregional control and progression-free survival in head and neck ACC without conferring overall survival advantage,likely reflecting the disease's characteristic indolent metastatic progression.These results establish>60 Gy as an optimal dose threshold for PORT in high-risk ACC management.

Objective:To investigate the clinical characteristics and prognosis of bloodstream infection caused by carbapenem-resistant Klebsiella pneumoniae (CRKP) in preterm infants, and to provide basis for early clinical diagnosis and infection control. Methods:The clinical data of infants with CRKP bloodstream infection admitted to the Preterm Infants Ward of Children′s Hospital Affiliated to Zhengzhou University from January 2015 to December 2022 were retrospectively analyzed. The risk factors for death in preterm infants caused by CRKP bloodstream infection were explored through multivariate logistic regression analysis, and the receiver operating characteristic (ROC) curve was used to analyze the clinical value of each factor on evaluating prognosis. The area under curves (AUC) of each factor in different ROC curve were compared by Delong′s test.Results:A total of 96 preterm infants with CRKP bloodstream infection were included, including 70 in the survival group and 26 in the death group. The first onset symptoms of CRKP bloodstream infection in preterm infants were persistent tachycardia (heart rate>180 per minute) (69 cases, 71.9%), fever (61 cases, 63.5%), and apnea (59 cases, 61.5%). There were 88(91.7%) cases of infection combined with septic shock, and 91(94.8%) cases required vasoactive drug support. Multivariate logistic regression analysis showed that the maximum vasoactive-inotropic score (VIS) within 48 hours of onset (odds ratio ( OR)=1.058, 95% confidence interval (95% CI) 1.022 to 1.095, P=0.001), concurrent purulent meningitis ( OR=8.029, 95% CI 1.344 to 47.972, P=0.022), and concurrent necrotizing enterocolitis (NEC) ( OR=10.881, 95% CI 1.566 to 75.580, P=0.016) were independent risk factors for death in preterm infants with CRKP bloodstream infection. The ROC curve showed that the AUCs for evaluating the prognosis of preterm infants with NEC and purulent meningitis were 0.784 and 0.711, respectively. The AUC for evaluating the prognosis of preterm infants with a maximum VIS ≥52.5 points within 48 hours of onset was 0.840, and the AUC for combining the three factors was 0.931. Compared with NEC and purulent meningitis, the AUC for combining factors was higher, the differences were statistically significant ( P=0.002, P<0.001). Conclusions:Preterm infants with CRKP bloodstream infection who have a maximum VIS ≥52.5 points within 48 hours of onset, with NEC and purulent meningitis have a higher risk of death.

OBJECTIVE To assess the clinical profile,prognosis,and risk factors of extrapulmonary metastasis in adenoid cystic carcinoma patients.METHODS A retrospective analysis was conducted on 126 patients diagnosed with adenoid cystic carcinoma at Beijing Tongren Hospital between January 2002 and December 2020.Of these patients,21 cases had metastases outside the lungs as their initial site of metastasis,while 105 cases had lungs as the initial site of distant metastasis.In addition,clinical data of patients diagnosed with adenoid cystic carcinoma from the Surveillance,Epidemiology,and End Results(SEER)database in the United States from 2010 to 2019 were analyzed for prognosis.RESULTS Univariate analysis showed that factors such as N stage,neurological symptoms,pathological subtype,grading,Ki67,neural invasion,and p63 status were associated with extrapulmonary metastasis(χ2=5.385,9.888,20.485,15.579,8.711,5.476,5.280;all P values<0.05).Multivariate logistic regression analysis indicated that N stage,pathological grading,and neurological symptoms were correlated with an increased risk of extrapulmonary metastasis.Survival analysis indicated lower cumulative survival and progression-free survival rates in the initial extrapulmonary metastasis group(both P values<0.05).CONCLUSION The initial metastasis site in adenoid cystic carcinoma is associated with multiple factors including N stage,pathological grading,and neurological symptoms.Patients displaying a solid subtype and those accompanied by high-grade transformation are more prone to extrapulmonary metastasis.Patients with extrapulmonary metastasis as their initial diagnosis typically exhibit poorer prognosis.

AIM:To investigate the inhibitory effect of the"decoction for soothing liver and removing stasis and toxicity(SGQYJDF)"on hepatocellular carcinoma(HCC)proliferation in nude mice by inducing ferroptosis via the tumor protein 53(p53)/solute carrier family 7 member 11(SLC7A11/xCT)/glutathione peroxidase 4(GPX4)pathway.METHODS:An ectopic subcutaneous tumor model was established by injecting SK-Hep-1 cells subcutaneously into the right axilla of nude mice.Upon formation of tumor,the mice were randomly divided into five groups(i.e.,control group,low-,medium-and high-dose SGQYJDF groups and medium-dose SGQYJDF plus Sorafenib group).Each group of mice was orally administered with the corresponding therapy for 14 consecutive days,during which the tumor size was observed regularly.At the end of treatment,the tumor growth inhibition rate was calculated based on tumor mass,and histopatho-logical changes were observed by HE staining.Then,the levels of malondialdehyde(MDA),glutathione(GSH)and fer-rous ions(Fe2+)were detected by colorimetric assays.The expression of the proliferation markers Ki-67 and GPX4 was de-tected by immunohistochemistry(IHC).The expression of p53 and xCT was detected by Immunofluorescence(IF).And the expression of p53,xCT and GPX4 was determined by Western blot.RESULTS:(1)SGQYJDF was found to dose-de-pendently decrease tumor volume(P＜0.01)and inhibit tumor mass growth(P＜0.01),and meanwhile,reduce the per-centage of Ki-67-positive cells(P＜0.01)and their proliferation ability in tumor tissues,as compared to the control group.(2)In terms of Ferroptosis-related indicators,SGQYJDF was found to dose-dependently increase the levels of Fe2+ and MDA but decrease the level of GSH in tumor tissues(P＜0.01),as compared to the control group.(3)In terms of protein expression,SGQYJDF was found to dose-dependently upregulate the expression of p53(P＜0.05)but inhibit the expres-sion of xCT(P＜0.05)and GPX4(P＜0.01).Notably,medium-dose SGQYJDF plus sorafenib showed a stronger effect than high-dose SGQYJDF.CONCLUSION:Our findings suggest that SGQYJDF can induce Ferroptosis to inhibit the proliferation of subcutaneously transplanted tumor tissues in nude mice by upregulating the expression of p53,and inhibit-ing the expression of xCT and GPX4.

Objective:To analyze the clinically acceptable and reproducible bladder and rectum volumes of prostate cancer patients during radiotherapy under bladder and bowel preparation, aiming to provide quantitative indicators for bowel and bladder preparation before and after radiotherapy.Methods:Clinical data of 275 prostate cancer patients with strict bladder and bowel preparation and completion of whole course radical radiotherapy at Sun Yat-sen University Cancer Center from April 2015 to December 2020 were retrospectively analyzed. Patients were scanned with cone beam CT (CBCT) before each treatment and the setup error was recorded. Sixty-six patients were selected by simple random sampling and the bladder and rectum on daily CBCT was outlined using MIM software. The relationship between the ratio of daily bladder or rectum volume to the planned bladder or rectum volume (relative value of volume) and setup error was analyzed. Quantitative data were expressed as mean±SD. Normally distributed data were analyzed by paired t-test while non-normally distributed data were assessed by Kruskal-Wallis test.Results:The bladder and rectum volume on planning CT were (370.87±110.04) ml and (59.94±25.07) ml of 275 patients. The bladder and rectum volumes on planning CT were (357.51±107.38) ml and (65.28±35.37) ml respectively of the 66 selected patients with 1611 sets of CBCT images. And the bladder and rectum volumes on daily CBCT were (258.96±120.23) ml and (59.95 ± 30.40) ml. The bladder volume of patients was decreased by 3.59 ml per day on average during the treatment and 0.37 ml for the rectum volume. According to the bladder volume on planning CT, all patients were divided into three groups: <250 ml, 250-450 ml and >450 ml groups. The relative value of volume in the 250-450 ml group during the course of radiotherapy was the smallest. And the setup error in the superior and inferior (SI) direction was (0.28±0.24) cm and (0.19±0.17) cm in the left and right (LR) direction, significantly lower than those in the other two groups (both P≤0.027). According to the rectum volume on planning CT, all patients were divided into four groups: <50 ml, 50-<80 ml, 80-120 ml and >120 ml groups. The <50 ml group had the smallest relative value of volume during radiotherapy, and the setup error in the SI direction was (0.26±0.22) cm and (0.24±0.22) cm in the anterior and posterior (AP) direction, significantly smaller than those in the other groups (both P≤0.003). The setup errors in the SI, LR, AP directions of the enrolled 66 patients were (0.30±0.25) cm, (0.20±0.18) cm and (0.28±0.27) cm, respectively. Among them, the relative value of bladder volume in the AP direction was (0.73±0.37) in the setup error <0.3 cm group, which was statistically different from those in the setup error 0.3-0.5 cm and >0.5 cm groups (both P<0.05). Conclusion:Under the bladder and bowel preparation before planning CT, the appropriate bladder and rectum volumes are in the range of 250-450 ml and <50 ml, which yields higher reproducibility and smaller setup error.

Objective:To evaluate the guidelines on discharged health education for solid organ transplant recipients at home and abroad, analyze the characteristics of each guideline, so as to provide reference for the development of health education for organ transplantation in China.Methods:This was a guideline quality evaluation study. The guidelines on health education for discharged recipients with solid organ transplantation were searched on the websites of clinical practice guidelines, the websites of transplantation professional associations, and Chinese and English databases at home and abroad through computers. The search time limit was from January 1, 2011 to January 1, 2022. The quality of guidelines that met the inclusion and exclusion criteria was evaluated using the Appraisal of Guidelines for Research and Evaluation Ⅱ (AGREE Ⅱ) .Results:A total of 8 guidelines were included. The overall quality evaluation of the guideline was Grade A ( n=2) and Grade B ( n=6) . In terms of standardized scores in six areas of quality evaluation, the average values of scope and purpose, stakeholder involvement, rigour of development, clarity and presentation, applicability as well as editorial independence were 87.62% (70.37%-100.00%) , 75.46% (44.44%-97.22%) , 66.54% (43.75%-93.06%) , 90.16% (77.78%-96.30%) , 48.18% (20.83%-68.75%) and 74.13% (8.30%-97.22%) , respectively. The recommendations of the guidelines covered 9 aspects: team composition and support services, drug management, infection prevention, complication management, diet and nutrition, daily life and social reintegration, contraception and reproduction, psychological guidance and follow-up review. Conclusions:The development of guidelines for different types of solid organ transplantation varies. The overall quality of the guidelines needs to be improved and the content needs to be refined. In the future, we can learn from foreign high-quality guidelines to carry out localization practice, so as to build a health education related guideline for discharge of solid organ transplantation that is suitable for China's national conditions as soon as possible.

OBJECTIVE To optimize the e xtraction technology of Guizhi shaoyao zhimu decoction （GSZD）. METHODS The contents of 9 components in GSZD were determined by HPLC ，such as ephedrine hydrochloride ，pseudoephedrine hydrochloride ， mangiferin，paeoniflorin，liquiritin，5-O-methylvisammioside，glycyrrhizic acid ，cinnamic acid ，6-gingerol. On the basis of single factor experiment ，taking material-liquid ratio ，extraction times and extraction time as inspection factors ，taking the contents of above 9 components and the yield of dry extract as evaluation indicators ，the analytic hierarchy process and entropy weight method were used to determine the composite weight of each index and calculate the comprehensive score ；the extraction technology parameters of GSZD were optimized by Box-Behnken response surface method ，and the validation tests were conducted. RESULTS The composite weight of the contents of ephedrine hydrochloride ，pseudoephedrine hydrochloride ，mangiferin，paeoniflorin， glycyrrhizin，5-O-methylvisa- midol ，glycyrrhizinate，cinnamic acid ，6-gingerol and the yield of dry extract were respectively 0.12，0.10，0.05，0.12，0.14，0.06，0.13，0.15，0.10，0.03. The optimal extraction technology of GSZD is that the ratio of material to liquid is 1 ∶ 14（g/mL），extraction is 2 times，and the extraction time is 3.0 h；average comprehensive score of the 3 verification tests was 95.879，and RSD was 0.50％（n＝3），the deviation from the predicted comprehensive score （94.328）was 1.64％. CONCLUSIONS In this study ，the optimal extraction technology of GSZD is determined.