ObjectiveTo elucidate the underlying mechanism through which Zhuluan decoction suppresses excessive autophagy in human ovarian granulosa cells （KGN） and ameliorates premature ovarian insufficiency （POI） via the phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）/mammalian target of rapamycin （mTOR） signaling pathway. MethodsThe optimal concentration of cyclophosphamide for inducing a POI model in KGN cells was identified via the cell counting kit-8 （CCK-8） assay. Subsequently， the impacts of varying concentrations of Zhuluan decoction-containing serum on the viability of the KGN cell model were assessed. After the optimal drug concentration was determined， KGN cells were categorized into the following groups： blank control （20% blank serum）， model （20% blank serum + 5 μmol·L^-1 cyclophosphamide）， Zhuluan decoction-containing serum （20% Zhuluan decoction-containing serum + 5 μmol·L^-1 cyclophosphamide）， autophagy inhibitor （20% blank serum + 5 μmol·L^-1 cyclophosphamide + 20 μmol·L^-1 chloroquine phosphate）， autophagy inhibitor + Zhuluan decoction-containing serum （20% Zhuluan decoction-containing serum + 5 μmol·L^-1 cyclophosphamide + 20 μmol·L^-1 chloroquine phosphate）， and estradiol valerate （20% estradiol valerate-containing serum + 5 μmol·L^-1 cyclophosphamide）. Following 48 hours of incubation， flow cytometry was utilized to measure the apoptosis rate of KGN cells in each group. Western blotting was employed to quantify the protein levels of PI3K， phosphorylated （p）-Akt， Akt， p-mTOR， and mTOR， along with the expression levels of autophagy-related proteins such as Beclin1， autophagy-related 5 homolog （ATG5）， and microtubule-associated protein 1 light chain 3 （LC3）， in each group. Additionally， monodansylcadaverine （MDC） staining was performed to evaluate the extent of autophagy in each group. ResultsIncubation of KGN cells with 5 μmol·L^-1 cyclophosphamide for 48 h successfully established a POI model， marked by a significant inhibition of KGN cell proliferation. Notably， the inhibitory effect of cyclophosphamide on KGN cell proliferation exhibited a positive correlation with its concentration. Zhuluan decoction-containing serum at 20% and 30% promoted cell proliferation and mitigated the inhibitory effect of cyclophosphamide on KGN cell proliferation， with comparable therapeutic efficacy observed at both concentrations. Compared with the blank control group， the model group displayed an elevated apoptosis rate （P<0.01）， reduced protein levels of PI3K， p-Akt， and p-mTOR （P<0.01）， increased protein levels of Beclin1， LC3， and ATG5 （P<0.01）， no significant alterations in the protein levels of Akt and mTOR， and an enhanced MDC autophagy fluorescence intensity （P<0.01）. In comparison to that the model group， the apoptosis rates in the blank control group， model group， Zhuluan decoction-containing serum group， autophagy inhibitor group， autophagy inhibitor + Zhuluan decoction-containing serum group， and estradiol valerate group all reduced （P<0.05， P<0.01）， with the most pronounced reduction observed in the autophagy inhibitor + Zhuluan decoction-containing serum group. The protein levels of PI3K， p-Akt， and p-mTOR were higher in other groups than in the model group （P<0.05， P<0.01）， being the highest in the autophagy inhibitor + Zhuluan decoctio-containing serum group （P<0.01）. The protein levels of Beclin1 and ATG5 were lower in other groups than in the model group （P<0.05， P<0.01）. The expression level of LC3 declined in the Zhuluan decoction-containing serum group and the estradiol valerate group （P<0.05， P<0.01）， while it decreased without statistical significance in the autophagy inhibitor group and the autophagy inhibitor + Zhuluan decoction-containing serum group. ConclusionZhuluan decoction may activate the PI3K/Akt/mTOR pathway to inhibit excessive autophagy and counteract the detrimental effects of cyclophosphamide on the KGN cell model， thus managing POI.

ObjectiveTo elucidate the underlying mechanism through which Zhuluan decoction suppresses excessive autophagy in human ovarian granulosa cells （KGN） and ameliorates premature ovarian insufficiency （POI） via the phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）/mammalian target of rapamycin （mTOR） signaling pathway. MethodsThe optimal concentration of cyclophosphamide for inducing a POI model in KGN cells was identified via the cell counting kit-8 （CCK-8） assay. Subsequently， the impacts of varying concentrations of Zhuluan decoction-containing serum on the viability of the KGN cell model were assessed. After the optimal drug concentration was determined， KGN cells were categorized into the following groups： blank control （20% blank serum）， model （20% blank serum + 5 μmol·L^-1 cyclophosphamide）， Zhuluan decoction-containing serum （20% Zhuluan decoction-containing serum + 5 μmol·L^-1 cyclophosphamide）， autophagy inhibitor （20% blank serum + 5 μmol·L^-1 cyclophosphamide + 20 μmol·L^-1 chloroquine phosphate）， autophagy inhibitor + Zhuluan decoction-containing serum （20% Zhuluan decoction-containing serum + 5 μmol·L^-1 cyclophosphamide + 20 μmol·L^-1 chloroquine phosphate）， and estradiol valerate （20% estradiol valerate-containing serum + 5 μmol·L^-1 cyclophosphamide）. Following 48 hours of incubation， flow cytometry was utilized to measure the apoptosis rate of KGN cells in each group. Western blotting was employed to quantify the protein levels of PI3K， phosphorylated （p）-Akt， Akt， p-mTOR， and mTOR， along with the expression levels of autophagy-related proteins such as Beclin1， autophagy-related 5 homolog （ATG5）， and microtubule-associated protein 1 light chain 3 （LC3）， in each group. Additionally， monodansylcadaverine （MDC） staining was performed to evaluate the extent of autophagy in each group. ResultsIncubation of KGN cells with 5 μmol·L^-1 cyclophosphamide for 48 h successfully established a POI model， marked by a significant inhibition of KGN cell proliferation. Notably， the inhibitory effect of cyclophosphamide on KGN cell proliferation exhibited a positive correlation with its concentration. Zhuluan decoction-containing serum at 20% and 30% promoted cell proliferation and mitigated the inhibitory effect of cyclophosphamide on KGN cell proliferation， with comparable therapeutic efficacy observed at both concentrations. Compared with the blank control group， the model group displayed an elevated apoptosis rate （P<0.01）， reduced protein levels of PI3K， p-Akt， and p-mTOR （P<0.01）， increased protein levels of Beclin1， LC3， and ATG5 （P<0.01）， no significant alterations in the protein levels of Akt and mTOR， and an enhanced MDC autophagy fluorescence intensity （P<0.01）. In comparison to that the model group， the apoptosis rates in the blank control group， model group， Zhuluan decoction-containing serum group， autophagy inhibitor group， autophagy inhibitor + Zhuluan decoction-containing serum group， and estradiol valerate group all reduced （P<0.05， P<0.01）， with the most pronounced reduction observed in the autophagy inhibitor + Zhuluan decoction-containing serum group. The protein levels of PI3K， p-Akt， and p-mTOR were higher in other groups than in the model group （P<0.05， P<0.01）， being the highest in the autophagy inhibitor + Zhuluan decoctio-containing serum group （P<0.01）. The protein levels of Beclin1 and ATG5 were lower in other groups than in the model group （P<0.05， P<0.01）. The expression level of LC3 declined in the Zhuluan decoction-containing serum group and the estradiol valerate group （P<0.05， P<0.01）， while it decreased without statistical significance in the autophagy inhibitor group and the autophagy inhibitor + Zhuluan decoction-containing serum group. ConclusionZhuluan decoction may activate the PI3K/Akt/mTOR pathway to inhibit excessive autophagy and counteract the detrimental effects of cyclophosphamide on the KGN cell model， thus managing POI.

Objective:To investigate the impact of early invasive arterial blood pressure (IBP) monitoring on survival and neurological outcomes in out-of-hospital cardiac arrest (OHCA) patients undergoing extracorporeal cardiopulmonary resuscitation (ECPR).Methods:This retrospective cohort study analyzed 44 OHCA patients receiving ECPR between January 2021 and January 2023. Patients were divided into: Early intervention group : IBP established within 3 min of ECMO initiation; Late intervention group : IBP established after ICU admission. Baseline characteristics, ECMO parameters, and clinical outcomes were compared. Multivariable logistic regression (adjusted for age, initial rhythm, etc.) and Spearman's correlation were used.Results:This study included a total of 44 patients treated with OHCA and ECPR, divided into an early intervention group of 23 cases and a late intervention group of 21 cases. The early intervention group showed significantly higher: Survival to discharge (43.5% vs. 9.5%, P<0.05), Good neurological recovery (CPC 1-2: 34.8% vs. 9.5%, P<0.05).Early intervention independently predicted survival (adjusted OR=18.84, 95% CI:1.97-179.98, P=0.01). Stratified analysis by pH (cutoff 7.0) demonstrated consistent benefits in both pH>7.0 ( aOR=0.392, 95% CI:0.106-0.678) and pH≤7.0 subgroups ( aOR=0.385, 95% CI: 0.075-0.695; interaction P=0.183). Early IBP positively correlated with CPC scores ( ρ=0.40, P=0.007). Conclusions:Early IBP monitoring significantly improves survival and neurological outcomes in OHCA-ECPR patients, supporting its integration into standardized protocols.

Objective:To investigate the transcriptional level expression of olfactomedin-like protein 2A (OLFML2A) in peripheral blood of patients with acute leukemia (AL) and its diagnostic and prognostic evaluation value.Methods:A retrospective cohort study was conducted. A total of 34 patients with AL (AL group) admitted to the Second People's Hospital of Lianyungang from January 2019 to March 2023 were selected, including 26 cases of acute myeloid leukemia (AML) (non-acute promyelocytic leukemia) and 8 cases of acute lymphoblastic leukemia (ALL). Another 15 healthy subjects who underwent the physical examination during the same period were selected as the healthy control group. Among 26 patients with AML, 18 were males and 8 were females. The median age [ M (IQR)] was 59 (9) years; 5 cases relapsed. Among 8 patients with ALL, 4 were males and 4 were females. The median age was 48 (12) years; 1 case relapsed. Real-time fluorescence quantitative polymerase chain reaction (qPCR) medthod was used to detect the relative expression level of OLFML2A mRNA in peripheral blood of each group. The relative expression levels of OLFML2A mRNA among the different patients stratified by clinicopathological factors were compared. Taking bone marrow smear cytology or bone marrow biopsy as the gold standard, receiver operating characteristic (ROC) curve was used to analyze the diagnostic effect of the relative expression level of OLFML2A mRNA on AML and ALL. According to the median relative expression level of OLFML2A mRNA in AL patients, the patients were divided into the high expression of OLFML2A mRNA (≥ the median) and the low expression of OLFML2A mRNA (< the median) groups. Progression-free survival (PFS) and overall survival (OS) of both groups were analyzed by using Kaplan-Meier curve. The log-rank test was used for comparison between groups. Results:The median relative expression level of OLFML2A mRNA in AL group was higher than that in the healthy control group [3.53 (8.19) vs. 0.27 (0.23)], and the difference was statistically significant ( Z = 4.38, P < 0.001). The median relative expression level of OLFML2A mRNA in AML group was higher than that in ALL group [4.92, (9.80) vs. 1.60 (4.35)], which were higher than that in the healthy control group; and the differences were statistically significant (all P < 0.05). There were statistically significant differences in the relative expression level of OLFML2A mRNA among AML patients with different prognosis risk stratification, fusion gene positive or not, whether there was chromosome abnormality, and whether the average daily hospitalization cost was < 2 500 yuan (all P < 0.05). There were statistically significant differences in the relative expression level of OLFML2A mRNA among ALL patients with different prognostic risk stratification, whether there was fusion gene and whether there was chromosome abnormality (all P < 0.05). ROC curve analysis showed that the area under the curve for the diagnosis of AML based on the relative expression level of OLFML2A mRNA was 0.936 (95% CI: 0.862-1.000), and the optimal cut-off value was 0.780; the area under the curve for the diagnosis ALL was 0.767 (95% CI: 0.535-0.998), and the optimal cut-off value was 0.558. Kaplan-Meier survival curve analysis showed that the 3-year PSF rate in AL patients with high expression of OLFML2A mRNA (17 cases) and low expression of OLFML2A mRNA was 20.5% and 30.6%, respectively, and PFS in AL patients with low expression of OLFML2A mRNA was superior to those with high expression, and the difference was statistically significant ( χ2 = 4.64, P = 0.031). The 3-year OS rates in AL patients with high and low expression of OLFML2A mRNA was 40.4% and 33.3%, respectively, and there was no statistically significant difference in OS between the 2 groups ( χ2 = 1.55, P = 0.213). Conclusions:The relative expression level of OLFML2A mRNA is high in AL patients, and it is more significant in AML patients. The level of OLFML2A gene has a certain value in the diagnostic and prognostic evaluation of AL.

Objective:To explore the relationship between Epstein-Barr virus (EBV) infection, hepatitis B virus (HBV) reactivation and clinical features and prognosis in HBV-infected non-Hodgkin lymphoma (NHL) patients and influencing factors of HBV reactivation.Methods:A retrospective cohort study was conducted. A total of 80 NHL patients with hepatitis B surface antigen (HBsAg) positive (which was defined as HBV positive) who were admitted to the Second People's Hospital of Lianyungang and Jiangsu Province Hospital from December 2012 to October 2022 were selected. All patients were divided into EBV-positive group and EBV-negative group according to EBV DNA results, and further grouped into the HBV reactivation group and the non-reactivation group according to whether HBV were reactivated after chemotherapy. The clinical characteristics of patients among groups were compared. Multivariate logistic regression model was used to analyze the factors influencing HBV reactivation. The Kaplan-Meier method was used to evaluate the progression-free survival (PFS) and overall survival (OS) of patients, and the log-rank test was used for inter-group comparison.Results:Among NHL patients with HBV positive, 27 cases (33.8%) were EBV-positive and 29 cases (36.3%) were HBV reactivation. Compared with the EBV-negative group, the proportion of patients with Ann Arbor stage Ⅲ-Ⅳ [92.6% (25/27) vs. 66.0% (35/53)], elevated β 2-microglobulin level [88.9% (24/27) vs. 62.3% (33/53)], bone marrow involvement [40.7% (11/27) vs. 15.1% (8/53)], and HBV reactivation [51.9% (14/27) vs. 28.3% (15/53)] was higher in the EBV-positive group, and the differences were statistically significant (all P<0.05). There were no statistically significant differences in the composition of patients stratified by age, gender, pathological type, B symptom, lactate dehydrogenase level, international prognostic index score, number of extranodal involvements, liver involvement, hepatitis outbreak, prophylactic anti-HBV therapy, hepatitis B surface antibody (HBsAb), rituximab therapy, and the last chemotherapy effects between the 2 groups (all P > 0.05). Compared with the HBV non-reactivation group, the proportion of patients undergoing hepatitis outbreak [48.3% (14/29) vs. 17.6% (9/51)], not receiving prophylactic anti-HBV therapy [65.5% (19/29) vs. 39.2% (20/51)], HBsAb negative [79.3% (23/29) vs. 21.6% (11/51)], EBV positive [48.3% (14/29) vs. 25.5% (13/51)], receiving rituximab [82.8% (24/29) vs. 60.8% (31/51)] was higher in the HBV reactivation group, and the differenves were statistically significant (all P < 0.05); while there were no statistically significant differences in the composition of patients stratified by the other clinical characteristics between the 2 groups (all P > 0.05). Multivariate logistic regression analysis showed that EBV-positivity was an independent risk factor for HBV reactivation after chemotherapy in NHL patients with HBsAg positive ( OR = 7.073, 95% CI: 1.613-31.010, P = 0.009), while HBsAb positive ( OR = 0.038, 95% CI: 0.008-0.186, P < 0.001) and preventive anti-HBV therapy ( OR = 0.172, 95% CI: 0.039-0.756, P = 0.020) were independent protective factors. The last follow-up was in December 2023 and the median follow-up time was 36.5 months. There were no statistically significant differences in PFS and OS between the EBV-positive group and the EBV-negative group, HBV reactivation group and the non-reactivation group (all P > 0.05). Conclusions:Among HBV-infected NHL patients, those with concurrent EBV infection have a more advanced clinical stage and are very prone to bone marrow invasion, and they also show a higher probability of HBV reactivation; HBV reactivation may be related to whether receiving preventive anti-HBV therapy and rituximab therapy. EBV infection may increase the risk of HBV reactivation in NHL patients; EBV infection and HBV reactivation may not be relevant to the prognosis of patients.

Acute T-lymphoblastic leukemia (T-ALL) is a hematopoietic malignancy, and in recent years, with the advancement of combined chemotherapy and hematopoietic stem cell transplantation, the prognosis of T-ALL has improved significantly, but for patients with primary drug resistance or relapsed/refractory disease the prognosis is still poor. The plant homeodomain finger 6 (PHF6) is a tumor suppressor protein, it plays a pivotal role in T cell differentiation, epigenetic regulation and oncogenic pathway synergy, and its mutations and deletions are commonly associated with the development of T-lymphocytic leukemia. However, the underlying mechanism of PHF6 in the pathogenesis of T-ALL remains unclear. This article reviews the structure, function and mechanism of action of PHF6 in T-ALL, the important coexisting genes associated with the progression of T-ALL, and the research progress in targeted therapy.

OBJECTIVE To investigate and analyze the current situation and problems of the advanced personnel en-gaging in the hospital-acquired infection control during their training period and explore the existing countermeas-ures and future development.METHODS The literatures regarding to the advanced study in China were retrieved from databases,the subjects of the literatures covered infection control-related advanced study practice,discipline construction,position competence,talent cultivation,scientific research innovation,professional title evaluation,laws,regulations and development plans.From Aug.2024 to Nov.2024,a questionnaire survey and face-to-face interviews were conducted among 36 advanced study personnel from 9 provinces of China who engaged in hos-pital-acquired infection control in the First Medical Center of Chinese PLA General Hospital.Eventually,36 ques-tionnaires were retrieved,all of which were valid with a questionnaire recovery rate of 100.00％.RESULTS Among the 36 advanced study personnel of hospital-acquired infection control,58.33％were medium-grade professional ti-tle;preventive medicine(41.67％),clinical medicine(25.00％)and nursing(16.67％)ranked the top 3 majors.The personnel engaged in the infection control for more than 6 years,and the duration of the advanced study was generally 3 or 6 months.In reality,the personnel faced the choices in terms of the purposes of further education,learning approaches and learning contents.The advanced study personnel also encountered the problems of challenges from promotion,improvement of position competency,integration with clinical training,supervision and practice,as well as physiological,psychological and family pressure.CONCLUSION Aiming at the problems that the advanced study personnel are generally concerned about,such as how to scientifically and effectively carry out hospital-acquired infection control advanced study and preset and solve the problems that may encounter,it is necessary to formulate targeted training programmes so as to provide bases and enlightenment for establishment of a long-term mechanism for advanced study of infection control in China.

Objective To identify the taste critical quality attribute and design and optimize the flavor-correcting formulation of the traditional Chinese medicine oral preparation Qingre Jiedu Oral Liquid,in order to improve its taste and enhance patient medication adherence.Methods The taste assignment method was employed to identify the taste critical quality attribute of Qingre Jiedu Oral Liquid.Based on human sensory evaluation and the standardized Euclidean distance in electronic tongue analysis,suitable types of corrigent were determined.Subsequently,under constraints such as maximum allowable dosage,solubility,and sweetness,the optimal taste formulation for the sugar-free intermediate of Qingre Jiedu Oral Liquid was determined using Box-Behnken experimental design combined with electronic tongue and human sensory evaluation results.The study was reviewed and approved by the Ethics Committee of Beijing University of Chinese Medicine(Ethics Approval Number 2020BZYLL0609).Results The quantitative score for bitter taste of Qingre Jiedu Oral Liquid accounted for 30.36%,confirming bitterness as the taste critical quality attribute requiring attention.The optimal taste formulation for the sugar-free intermediate of Qingre Jiedu Oral Liquid was determined to be 120 mg·mL?1 erythritol,12 mg·mL?1 acesulfame potassium,and 2.4 mg·mL?1 stevioside.This formulation achieved an 11.75-point improvement in sensory evaluation scores compared to the original commercially available oral liquid.Conclusion This study successfully improved the taste of Qingre Jiedu Oral Liquid and established a comprehensive strategy for flavor-correcting formulation optimization,including a method for identifying taste critical quality attribute.This strategy provides a referential paradigm for palatability enhancement of similar traditional Chinese medicine oral preparations,laying a crucial technical foundation for elevating the clinical value of Chinese herbal medicines and promoting the high-quality development of traditional Chinese medicine(TCM).

OBJECTIVE To investigate and analyze the current situation and problems of the advanced personnel en-gaging in the hospital-acquired infection control during their training period and explore the existing countermeas-ures and future development.METHODS The literatures regarding to the advanced study in China were retrieved from databases,the subjects of the literatures covered infection control-related advanced study practice,discipline construction,position competence,talent cultivation,scientific research innovation,professional title evaluation,laws,regulations and development plans.From Aug.2024 to Nov.2024,a questionnaire survey and face-to-face interviews were conducted among 36 advanced study personnel from 9 provinces of China who engaged in hos-pital-acquired infection control in the First Medical Center of Chinese PLA General Hospital.Eventually,36 ques-tionnaires were retrieved,all of which were valid with a questionnaire recovery rate of 100.00％.RESULTS Among the 36 advanced study personnel of hospital-acquired infection control,58.33％were medium-grade professional ti-tle;preventive medicine(41.67％),clinical medicine(25.00％)and nursing(16.67％)ranked the top 3 majors.The personnel engaged in the infection control for more than 6 years,and the duration of the advanced study was generally 3 or 6 months.In reality,the personnel faced the choices in terms of the purposes of further education,learning approaches and learning contents.The advanced study personnel also encountered the problems of challenges from promotion,improvement of position competency,integration with clinical training,supervision and practice,as well as physiological,psychological and family pressure.CONCLUSION Aiming at the problems that the advanced study personnel are generally concerned about,such as how to scientifically and effectively carry out hospital-acquired infection control advanced study and preset and solve the problems that may encounter,it is necessary to formulate targeted training programmes so as to provide bases and enlightenment for establishment of a long-term mechanism for advanced study of infection control in China.

Objective To identify the taste critical quality attribute and design and optimize the flavor-correcting formulation of the traditional Chinese medicine oral preparation Qingre Jiedu Oral Liquid,in order to improve its taste and enhance patient medication adherence.Methods The taste assignment method was employed to identify the taste critical quality attribute of Qingre Jiedu Oral Liquid.Based on human sensory evaluation and the standardized Euclidean distance in electronic tongue analysis,suitable types of corrigent were determined.Subsequently,under constraints such as maximum allowable dosage,solubility,and sweetness,the optimal taste formulation for the sugar-free intermediate of Qingre Jiedu Oral Liquid was determined using Box-Behnken experimental design combined with electronic tongue and human sensory evaluation results.The study was reviewed and approved by the Ethics Committee of Beijing University of Chinese Medicine(Ethics Approval Number 2020BZYLL0609).Results The quantitative score for bitter taste of Qingre Jiedu Oral Liquid accounted for 30.36%,confirming bitterness as the taste critical quality attribute requiring attention.The optimal taste formulation for the sugar-free intermediate of Qingre Jiedu Oral Liquid was determined to be 120 mg·mL?1 erythritol,12 mg·mL?1 acesulfame potassium,and 2.4 mg·mL?1 stevioside.This formulation achieved an 11.75-point improvement in sensory evaluation scores compared to the original commercially available oral liquid.Conclusion This study successfully improved the taste of Qingre Jiedu Oral Liquid and established a comprehensive strategy for flavor-correcting formulation optimization,including a method for identifying taste critical quality attribute.This strategy provides a referential paradigm for palatability enhancement of similar traditional Chinese medicine oral preparations,laying a crucial technical foundation for elevating the clinical value of Chinese herbal medicines and promoting the high-quality development of traditional Chinese medicine(TCM).