Objective : To explore the role of lysophosphatidylcholine acyltransferase 3(LPCAT3) in Erastin-induced ferroptosis of acute myeloid leukemia(AML) cells and its related molecular regulatory mechanisms. Methods : Tetrazolium salt(MTS) method was used to detect the sensitivity of different AML cells to the classic ferroptosis inducer Erastin, real time quantitative polymerase chain reaction(qPCR) was used to detect the basal expression level ofLPCAT3mRNA, and the correlation between them was analyzed. Lentivirus-mediatedLPCAT3overexpression AML cell lines(OE group) and negative control lines(NC group) were constructed. After Erastin intervention, MTS, flow cytometry, and micromethods were used to detect cell viability, lipid reactive oxygen species(ROS), and Malondialdehyde(MDA), respectively. qPCR and Western blot were used to detect unfolded protein response(UPR) classic pathway signaling molecules(PERK, ATF4, GRP78, etc.) expression levels. The above ferroptosis-related indicators were detected after combined intervention with the UPR inhibitor 4-phenylbutyric acid(4-PBA), and the regulatory relationship was analyzed. Results : Four different types of AML cells had different sensitivities to ferroptosis, among which K562 cells were relatively insensitive. The IC50of the four types of AML cells to Erastin was negatively correlated with the expression level ofLPCAT3(r=-0.919,P<0.001). After Erastin intervention, the cell viability of K562 cells in the OE group was significantly inhibited by Erastin compared with the NC group(P<0.001), and the levels of lipid ROS and MDA increased(P<0.001). The results of qPCR and Western blot showed that, compared with the NC group, the mRNA and protein expression of UPR classic pathway moleculesPERK,ATF4, andGRP78mRNA and protein increased in the OE group(P<0.01). After inhibiting the UPR pathway by 4-PBA, the viability of K562 cells decreased(P<0.01), and lipid ROS and MDA levels increased(P<0.01) compared with the uninhibited state. Conclusion Overexpression ofLPCAT3can promote ferroptosis in K562 cells, and this process is negatively regulated by the classical UPR pathway PERK/ATF.

[Objective] To explore the feasibility of using whole blood as a source of NK cells for allogeneic CAR NK cell therapy and activated NK cell reinfusion therapy, and initially construct a technical system for the separation and purification of NK cells from whole blood. [Methods] All peripheral blood mononuclear cells (PBMCs) were enriched from 400 mL of whole blood by manual separation and machine separation, respectively. The erythrocyte loss rate, PBMCs number, NK cell purity of the two methods were compared. NK cells were sorted from PBMCs by three separation and enrichment methods as immunomagnetic bead negative selection method, platelet lysate culture expansion and PERCOLL density gradient separation method, and the purity and yield of NK cells, the activity of NK cells and the tumor-killing ability of the three separation and enrichment methods were compared. [Results] The proportion of NK cells in the lymphocyte population was higher in the manual separation method than in the machine separation method[(13.16±5.16)% vs (8.56±3.92)%, P<0.05]; the number PBMCs was lower in the manual separation method than in the machine separation method[(4.09±1.80)×10⁸vs (6.49±2.16)×10⁸, P<0.05], and there was no difference in the red blood cell loss between the two methods (P>0.05). The purity of NK cells isolated and enriched from PBMCs by manual separation method using immunomagnetic was (96.77±2.31)%; the yield was (56.27±10.47)%; the inhibition of tumor proliferation was (38.67±14.05)%; and the tumor killing rate was (19.90±8.05)%. The purity of NK cells isolated and enriched from PBMCs by manual separation method using platelet lysis culture expansion method was the highest at day 7, which was (54.84±15.80)%; the cell expansion multiple could reach 16.92±6.28 at day 7; the in vitro tumor killing rate of NK cells was (15.83±5.5)%; the tumor inhibition rate was (44.33±13.5)%; and there was no difference in the toxicity and activity of NK cells between the two methods (P>0.05). The purity of NK cells isolated and enriched by PERCOLL density gradient separation method was (15.83±5.82)%, and the yield was (14±6.25)%, which was significantly lower than the other two methods. [Conclusion] PBMCs isolated from whole blood by manual separation and NK cells enriched by negative selection with immunomagnetic beads have the potential to provide NK cell materials for CAR-NK cell therapy, and NK cells enriched by platelet lysate-conditioned medium have the potential to provide NK cells for large-scale NK cell activation reinfusion therapy.

Objective:To investigate the risk factors associated with post-prematurity respiratory disease (PPRD) in very preterm infants.Methods:A prospective cohort study was conducted, enrolling 369 very preterm infants who were admitted to the neonatal intensive care unit of Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, within one week of birth from January 2019 to June 2023. Data on maternal and infant clinical characteristics, neonatal morbidities, and treatments during hospitalization were collected. The very preterm infants were divided into 2 groups based on whether they developed PPRD. Continuous variables were compared using Mann-Whitney U test, while categorical variables were compared using χ2 tests or continuity correction χ2 test. Multivariate Logistic regression analysis was used to identify the independent risk factors for PPRD in very preterm infants. Results:Among the 369 very preterm infants, 217 cases(58.8%) were male, with a gestational age of 30 (28, 31) weeks at birth and a birth weight of 1 320 (1 085, 1 590) g. Of these, 116 cases (31.4%) developed PPRD, while 253 cases (68.6%) did not. The very preterm infants in the PPRD group had a lower gestational age and lower birth weight (both, P<0.001). The PPRD group also had a higher proportion of males, lower Apgar scores at the 1 th minute after birth and the 5 th minutes after birth, a higher rate of born via cesarean delivery, and a higher incidence of bronchopulmonary dysplasia, more pulmonary surfactant treatment, longer durations of mechanical ventilation, longer total oxygen therapy, and lower Z-score for weight at discharge (all P<0.05). Multivariate Logistic regression analysis showed that gestational age ( OR=0.85, 95% CI 0.73-0.99, P=0.037), born via cesarean delivery ( OR=2.23, 95% CI 1.21-4.10, P=0.010), a duration of mechanical ventilation ≥7 days ( OR=2.51, 95% CI 1.43-4.39, P=0.001), and a Z-score for weight at discharge ( OR=0.82, 95% CI 0.67-0.99, P=0.040) were all independent risk factors for PPRD in very preterm infants. Conclusion:Very preterm infants with a small gestational age, born via cesarean section, mechanical ventilation ≥7 days, and a low Z-score for weight at discharge should be closely monitored for PPRD, and provided with standardized respiratory management after discharge.

OBJECTIVE To investigate the reference range of steady-state trough concentrations in depression patients taking paroxetine. METHODS Therapeutic drug monitoring data of 890 depression inpatients treated with paroxetine in our hospital from January 1， 2023 to December 31， 2024 were retrospectively collected. Univariate analysis and multiple linear regression analysis were employed to explore the influencing factors of the steady-state trough concentration of paroxetine， as well as the correlation between concentration and efficacy and adverse reactions. The reference range of steady-state trough concentration was obtained by receiver operating characteristic（ROC） curve method. RESULTS Patients with a greater degree of improvement in therapeutic efficacy exhibited higher steady-state trough concentrations of paroxetine （P＜0.000 1）. The steady-state trough concentrations of paroxetine and the ratio of paroxetine steady-state trough concentration to dose （C/D ratio） were significantly lower in male patients and those weighing 60-80 kg compared to female patients and those weighing＜60 kg， respectively （P＜0.05 or P＜0.000 1）. The steady-state trough concentration， C/D ratio， dosage， and concomitant medication all showed a positive correlation with therapeutic efficacy （P＜0.05 or P＜0.000 1）. Both the steady-state trough concentration and C/D ratio were correlated with liver function impairment， and the C/D ratio was also correlated with urinary retention （P＜0.05 or P＜0.000 1）. The critical threshold for the effective concentration of paroxetine was 56.31 ng/mL in the overall population， 56.42 ng/mL in males， 44.91 ng/mL in females， and 198.90 ng/mL in patients experiencing adverse reactions. CONCLUSIONS The reference range for the steady-state trough concentration of paroxetine in the overall population is 56.31-198.90 ng/mL； for male patients， it is 56.42-198.90 ng/mL， and for female patients， it is 44.91-198.90 ng/mL. Dosage of paroxetine should be reduced as appropriate for female patients and patients with low body weight or abnormal liver function.

Objective To monitor the antifungal resistance of clinical isolates of Candida spp.in the East China region.Methods MALDI-TOF MS or molecular methods were used to re-identify the strains collected from January 2018 to December 2022.Antifungal susceptibility testing was performed using the broth microdilution method.The susceptibility test results were interpreted according to the breakpoints of 2022 Clinical and Laboratory Standards Institute(CLSI)documents M27 M44s-Ed3 and M57s-Ed4.Results A total of 3 026 strains of Candida were collected,65.33％of which were isolated from sterile body sites,mainly from blood(38.86％)and pleural effusion/ascites(10.21％).The predominant species of Candida were Candida albicans(44.51％),followed by Candida parapsilosis complex(19.46％),Candida tropicalis(13.98％),Candida glabrata(10.34％),and other Candida species(0.79％).Candida albicans showed overall high susceptibility rates to the 10 antifungal drugs tested(the lowest rate being 93.62％).Only 2.97％of the strains showed dose-dependent susceptibility(SDD)to fluconazole.Candida parapsilosis complex had a SDD rate of 2.61％and a resistance rate of 9.42％to fluconazole,and susceptibility rates above 90％to other drugs.Candida glabrata had a SDD rate of 92.01％and a resistance rate of 7.99％to fluconazole,resistance rates of 32.27％and 48.24％to posaconazole and voriconazole non-wild-type strains(NWT),respectively,and susceptibility rates above 90％to other drugs.Candida tropicalis had resistance rates of 29.55％and 26.24％to fluconazole and voriconazole,respectively,resistance rates of 76.60％and 21.99％to posaconazole and echinocandins non-wild-type strains(NWT),and a resistance rate of 2.36％to echinocandins.Conclusions The prevalence and species distribution of Candida spp.in the East China region are consistent with previous domestic and international reports.Candida glabrata exhibits certain degree of resistance to fluconazole,while Candida tropicalis demonstrates higher resistance to triazole drugs.Additionally,echinocandins resistance has emerged in Candida albicans,Candida glabrata,Candida tropicalis,and Candida parapsilosis.

Objective:To investigate the effect and mechanism of miR-217 targeted regulation of extracellular signal-regulated kinase 2(ERK2)expression on activity and immune escape of non-small cell lung cancer cells(NSCLC).Methods:qRT-PCR was used to detect expression levels of miR-217 and ERK2 mRNA in NSCLC tissues,adjacent tissues,and HLF-1,A549 and HCC827 cell lines.Analyzed prognosis and survival status of NSCLC patients with different miR-217 expression level.Bioinformatics and dual luciferase gene reporting experiments were used to analyze the targeting relationship between miR-217 and ERK2.Cultivated NSCLC A549 cells and divided them into NC group,miR-217 inhibitor group,miR-217 mimic group,miR-217 mimic+ERK2 NC group and miR-217 mimic+ERK2 group.Except for the NC group without any treatment,all other groups were transfected with corresponding plasmids to analyze the proliferation activity and immune escape status of A549 cells in each group,and clarified the mechanism of action.Results:Compared with adjacent tissues,expression of miR-217 in NSCLC tissue was decreased,while expression of ERK2 mRNA was increased(P<0.05).Compared with human normal lung fibroblast HLF-1 cell lines,expression of miR-217 in NSCLC cell lines A549 and HCC827 were decreased,while expression of ERK2 mRNA was increased(P<0.05).Analysis of the relationship be-tween miR-217 and prognosis of NSCLC patients based on Kaplan-Meier Plotter database showed that low expression of miR-217 was associated with poor prognosis of patients(HR=0.90,P=0.033).Dual fluorescein reporter genes showed matching sequences between the 3'UTR regions of miR-217 and ERK2.miR-217 mimic fragment could inhibit ERK2-WT signal,but had no effect on ERK2-MUT.Compared with NC group,cell proliferation activity,PD-L1 and PD-L2 mRNA expression levels of miR-217 inhibitor group were in-creased,while CD8+T cell activity was decreased,and cell proliferation activity,PD-L1 and PD-L2 mRNA expression levels of miR-217 mimic group were decreased,while CD8+T cell activity was increased(P<0.05).Compared with miR-217 mimic group,cell pro-liferation activity,CD8+T cell activity,PD-L1 and PD-L2 mRNA expression levels of miR-217 mimic+ERK2 NC group had no signifi-cant changes(P>0.05),cell proliferation activity,PD-L1 and PD-L2 mRNA expression levels of miR-217 mimic+ERK2 group were increased,while CD8+T cell activity was decreased(P<0.05).Conclusion:Overexpression of miR-217 can reduce the activity of NSCLC cell A549,inhibit the expression of PD-L1,activate CD8+T cells in tumor microenvironment,and then inhibit immune es-cape,which may play a role by targeting ERK2.

Objectives:To explore the feasibility of using hemodynamic phenotypes to guide antihypertensive treatment medication. Methods:This study prospectively included 100 hypertensive patients who received outpatient treatment at Haidian Hospital in Beijing from January 2021 to December 2021.Evaluation of blood pressure was conducted using laboratory and home blood pressure measurements,impedance cardiogram(ICG)detection was performed.Following hemodynamic phenotypes and therapeutic phenotypes were established:hyperkinetic phenotype(increased heart rate)using β-blockers,large artery phenotype(increased large artery resistance index)using calcium antagonists,peripheral vascular phenotype(increased peripheral vascular resistance index)using renin angiotensin system inhibitors,and high-volume phenotype(increased blood volume saturation)using diuretics.Patients were randomly divided into ICG group(n=50,medication treatment based on hemodynamic characteristics)and control group(n=50,treatment based on hypertension guidelines and clinical experience).Patients were followed up for 8 weeks and the blood pressure reduction amplitude and compliance rate were compare between the two groups. Results:There was no statistically significant difference in baseline data such as sex,age,height,weight,and hemodynamic parameters between the two groups(all P＞0.05).There was no statistically significant difference in the types of baseline medication between the two groups(P＞0.05).After medication adjustment,the types of medication increased,but the difference between the two groups was still not statistically significant(P＞0.05).Clinic blood pressure:After 8 weeks,decreases in systolic([8.38±27.78]mmHg,1 mmHg=0.133 kPa)and diastolic([3.94±18.15]mmHg)blood pressure were greater in the ICG group as compared to the control group(both P＜0.05).The blood pressure compliance rate(＜140/90 mmHg)was higher in the ICG group than that in the control group(66.0%vs.42.0%,P＜0.05).Family blood pressure:after 8 weeks,the reduction in systolic([8.22±21.31]mmHg,P＜0.01)and diastolic([4.76±13.88]mmHg,P＜0.05)blood pressure was greater in the ICG group compared to the control group.The blood pressure compliance rate(＜135/85 mmHg)was higher in the ICG group than that in the control group(70.0%vs.48.0%,P＜0.05).Changes in corresponding hemodynamic parameters before and after two different antihypertensive drugs:the heart rate,arterial resistance index,peripheral vascular resistance index,and blood volume saturation of the ICG group all significantly decreased compared to baseline(all P＜0.05).The control group showed a significant decrease in peripheral vascular resistance index compared to baseline(P＜0.05),while there was no statistically significant difference in heart rate,large artery resistance index,and blood volume saturation compared to baseline(all P＞0.05). Conclusions:By using impedance cardiogram to assess hemodynamic phenotypes and accurately guide the selection of antihypertensive drugs based on hemodynamic phenotypes,it is possible to more effectively lower blood pressure and improve blood pressure compliance.

Allogeneic hematopoietic stem cell transplantation is an effective method for treating various hematological diseases. Membranous nephropathy is the second leading cause of novo glomerular disease after transplantation. Protocadherin FAT1 is a target antigen for diagnosing membranous nephropathy, primarily related to hematopoietic stem cell transplantation. Here we report a rare case of membranous nephropathy after hematopoietic stem cell transplantation. The target antigen of membranous nephropathy was confirmed to be FAT1 by pathology combined with mass spectrometry analysis. The nephrotic syndrome achieved partial remission after corticosteroid combined with cyclosporine treatment, and the patient's condition was stable during follow-up.

This article reports two cases of proteinuria caused by anti-vascular endothelial growth factor (VEGF) drugs in the treatment of liver cancer and ovarian cancer, respectively. Hypertension, massive albuminuria and renal insufficiency occurred in the patient with liver cancer during the use of toripalimab/camrelizumab combined with lenvatinib and regorafenib, and urinary protein was 4.02 g/24 h. The patient with ovarian cancer developed hypertension, proteinuria and 1.13 g/24 h urinary protein during the use of bevacizumab. Renal biopsy showed thrombotic microangiopathy in the glomeruli of both patients. Anti-VEGF drugs-associated glomerular microangiopathy was considered. The patient with liver cancer had no improvement after halving the dose of lenvatinib, and the effect was not good after treatment with low-dose corticosteroids combined with renin-angiotensin-aldosterone system inhibitors. After stopping anti-VEGF drugs, the proteinuria in both patients turned negative.

Objective To analyze the management and countermeasures of adverse nursing events in a municipal tertiary hospital in Guangxi Zhuang Autonomous Region.Methods The adverse nursing events in 2023 in a municipal tertiary hospital in Guangxi were retrospectively analyzed.Logistic regression analysis was utilized to identify the causes of these adverse events and propose suitable countermeasures.Results In 2023,a total of 121 adverse nursing events were recorded,with falls being the most prominent(42 cases,accounting for 34.71％),followed by drug extravasation,drug safety incidents and unplanned ex-tubation,accounting for 13.22％,9.92％,and 7.44％respectively.Logistic regression analysis revealed that factors such as age ≥70 years,coexisting diseases,Basel Index score ≤40,pain score ≥4,tube slip risk assessment ≥10,RASS sedation score ≥3,NGASR score ≥9,lost risk assessment score ≥ 7,GCS Coma Scale score ≤8,Stress Injury Assessment Scale score ≤10,and Morse score ≥45 were associated with the risk of adverse nursing events.In addition,inadequate awareness of nursing risks among nursing staff,ineffective patient health education methods,and non-compliance to nursing protocols were identified as primary causes of adverse nursing events.Conclusion Advanced age,combined with other diseases,and hospitali-zation scale assessment results are critical factors for adverse nursing events.Lack of nursing responsibility and risk awareness among nursing staff,inadequate system implementation,and irregular operations can increase the risk.Hospitals should optimize management systems and work processes,enhance nursing staff training,and develop targeted preventive measures for adverse nursing events to improve the quality of nursing management and reduce the risk of adverse nursing events.