BACKGROUND:Solute carrier family 1 member 5(SLC1A5)plays a potential role in a variety of diseases,but the exact mechanism of action is unclear.The construction of stable SLC1A5 overexpression and knockdown cell models can provide a powerful experimental tool for in-depth study of the exact role and mechanism of SLC1A5 in diseases and the discovery of potential therapeutic targets. OBJECTIVE:To construct lentiviral vectors for overexpression and knockdown of mouse SLC1A5 and establish stable transfected RAW264.7 cell lines,so as to provide an experimental foundation for further investigation of the role of SLC1A5 in inflammation. METHODS:Primers were designed and synthesized based on the SLC1A5 gene sequence,and the gene segment was amplified using polymerase chain reaction.Subsequently,the target gene segment was directionally inserted into the GV492 vector plasmid,which had been digested with AgeI/NheI enzymes,to construct recombinant lentiviral plasmids.Positive clones were further selected,and their sequences were confirmed.The pHelper1.0 plasmid vector and pHelper2.0 plasmid vector,along with the target plasmid vector,was co-cultured with 293T cells for transfection,resulting in the production and titration of lentiviral stocks.Furthermore,RAW264.7 cells were cultured in vitro,and the working concentration of puromycin was determined.Lentiviruses were separately co-cultured with RAW264.7 cells,and transfection efficiency was determined by measuring fluorescence intensity.Stable transfected cells were selected using puromycin,and real-time fluorescence quantitative PCR and western blot assay were employed to assess the gene and protein expression levels of SLC1A5 in stably transfected cell lines. RESULTS AND CONCLUSION:(1)Sequencing results indicated a perfect match between the sequencing and target sequences,confirming the successful construction of recombinant lentiviral vectors.(2)The titer for the overexpression SLC1A5 lentivirus was 1×109 TU/mL,while the titer for the knockdown SLC1A5 lentivirus was 3×109 TU/mL.(3)The working concentration of puromycin for RAW264.7 cells was determined to be 3 μg/mL.(4)The optimal conditions for transfecting RAW264.7 cells with overexpression/knockdown expression of SLC1A5 lentivirus involved the use of HiTransG P transfection enhancer with a multiplicity of infection value of 50.(5)A significant upregulation of the gene and protein expression levels of SLC1A5 was detected in cell lines stably overexpressing SLC1A5,while gene and protein expression levels of SLC1A5 were significantly decreased in the knockdown stable cell lines.These findings indicate that lentiviral vectors for mouse SLC1A5 overexpression and knockdown have been successfully constructed and a stably transfected RAW264.7 cell line has been obtained.

Objective:To observe any effect of oscillating positive expiratory pressure training on the airway clearing ability of postoperative esophageal cancer patients after neoadjuvant chemotherapy.Methods:Forty postoperative esophageal cancer patients undergoing neoadjuvant chemotherapy were enrolled and randomized into a control group and an experimental group, each of 20. Both groups received conventional postoperative rehabilitation starting on the first postoperative day, while the experimental group additionally underwent oscillating positive expiratory pressure training (3 sets/day, 30 breaths/set) for five consecutive days. Peak cough flow was measured using a peak flow meter before and 1, 3 and 5 days after the operation. Forced expiratory volume in the first second (FEV1), forced vital capacity (FVC), and peak expiratory flow (PEF) were also measured on the 5th day after the operation using spirometry. Any postoperative pulmonary complications were recorded.Results:On the 1st day after the operation, peak cough flow had decreased significantly in both groups compared to preoperative levels. However, it had increased significantly on days 3 and 5 in both groups, with the average increase in the experimental group significantly greater than in the control group. On day 5 the average FEV1, forced vital capacity and PEF in both groups were significantly lower than the preoperative values, while the experimental group demonstrated significantly higher average FEV1s (2.22±0.51L) and PEFs (5.09±1.26L/s) compared to the control group.Conclusions:Early postoperative oscillating positive expiratory pressure training can improve the peak cough flow and airway clearing ability of esophageal cancer patients after neoadjuvant chemotherapy, promoting the recovery of their lung function.

Objective To investigate the relationship between serum trimethylamine oxide(TMAO),neurofilament light chain protein(NfL),peroxisome proliferator-activated receptor γ coactivator-1α(PGC-1α)expression levels and short-term prognosis in aneurysmal subarachnoid hemorrhage(aSAH)patients.Method A total of 125 aSAH patients(aSAH group)and 125 healthy volunteers in the same period(control group)who were admitted in heji hospital affiliated to Changzhi Medical College from March 2020 to June 2023 were selected.The serum expression levels of TMAO,NfL and PGC-1α were compared between control group and aSAH group.The aSAH patients were followed up for 6 months after discharge.Their prognosis were evaluated using Glasgow Outcome Scale(GOS)and they were further divided into good prognosis and poor prognosis groups according to the GOS results.The serum expression levels of TMAO,NfL and PGC-1α were compared between the two groups.The poor prognosis influencing factors were analyzed by multivariate Logistic regression analysis,the serum TMAO,NfL and PGC-1α value in predicting poor prognosis were analyzed by receiver operating characteristic(ROC)curve.Result The expression levels of serum TMAO and PGC-1 α in the aSAH group were(2.63±0.36)μmol/L and(0.51±0.13)ng/mL,respectively,which were lower than those in the control group(3.18±0.57)μmol/L and(0.81±0.16)ng/mL(P<0.05).The expression level of serum NfL was significantly higher in the aSAH group(64.48±14.35 pg/mL)than in the control group(28.36±8.82 pg/mL)(P<0.05).Compared with the good prognosis group whose serum levels of TMAO and PGC-1 α were(2.80±0.80)μmol/L and(0.58±0.16)ng/mL,respectively,the poor prognosis group had significantly lower serum TMAO[(2.29±0.63)μmol/L]and PGC-1 α[(0.36±0.12)ng/mL](P<0.05).In contrast,poor prognosis group had a significantly higher level of NfL(76.70±15.61)pg/mL compared to good prognosis group(58.52±10.52)pg/mL(P<0.05).The proportion of patients with hypertension,patients with diabetes,patients with large or giant aneurysms,patients with Hunt Hess grade Ⅲ-Ⅳ,patients with onset to hospital time>12 h,and the level of C-reactive protein(CRP)were higher in the poor prognosis group than in the good prognosis group(P<0.05).Hunt Hess grade Ⅲ-Ⅳ,elevated serum CRP and NfL were independent risk factors for poor prognosis in aSAH patients(P<0.05),while elevated TMAO and PGC-1 α were protective factors(P<0.05).The area under the curve(AUC)of serum TMAO,NfL,PGC-1 α,and their combined prediction of poor prognosis in aSAH patients were 0.726,0.830,0.862,and 0.956,respectively.The AUC of the combined detection was greater than that of each indicator detected separately.Conclusion Serum TMAO and PGC-1α are lowly expressed in aSAH patients,and serum NfL is highly expressed,which are related to the occurrence of short-term poor prognosis,the combined detection of the three indicators has a high predictive value for short-term poor prognosis in aSAH patients.

Objective:To explore the lived experiences of family members of terminal ICU patients regarding their humanistic care needs and provide theoretical foundations for developing nursing care plans tailored to their needs.Methods:This study was a descriptive qualitative study. From April to December 2023, 16 family members of terminally ill ICU patients in Shanghai East Hospital, Tongji University were selected for semi-structured interviews using purposive sampling method, and the interview data were qualitatively analyzed using Colaizzi 7-step analysis.Results:The humanistic care needs of family members of terminally ill ICU patients can be categorized into five themes, namely, the need to know the condition at the first time; the need to participate in treatment and decision-making; the need to respect the wishes of terminally ill patients; the need for psychological care; and the need for social support.Conclusions:The humanistic care needs of family members of terminal ICU patients remain largely unmet. Nursing professionals should consider these needs and preferences and provide family members with professional guidance to help them establish positive coping mechanisms.

Objective This study aims to observe the impact of warm moxibustion on menstrual pain in college students with primary dysmenorrhea and explore its potential mechanism.Methods College students with primary dysmenorrhea were recruited and treated with warm moxibustion at Shenque acupoint for three consecutive menstrual cycles.Healthy subjects were also recruited for comparison.Pain scale,uterine artery hemodynamics,and related inflammatory factors were assessed before and after treatment.Results ①The results of scale study showed that the severity and duration of dysmenorrhea were gradually alleviated with the prolongation of treatment time through the analysis of variance of repeated measurements of the total scores of McGill pain inquiry scale and CMSS dysmenorrhea symptom scale before and after 3 treatments.The results of variance analysis and pairwise comparison of repeated measurements of PRI,VAS and PPI of McGill pain inquiry scale before and after 3 times treatment in warm moxibustion group also showed that each index decreased gradually with the prolongation of treatment time.The comparison of the scores of each item of CMSS scale showed that the severity and duration of low back before treatment were significantly different from those in the healthy group(P<0.001),but the difference was weakened after the third treatment.The severity of vomiting,the duration of vomiting,the severity of diarrhea and the duration of diarrhea were significantly different from those in the healthy group before treatment(P<0.001),but they were still higher than those in the healthy group after the third treatment.but the difference was not statistically significant.②Prior to treatment,PD college students exhibited significantly higher S/D and PI values on both sides compared to healthy subjects,with a statistically significant difference observed for PI on the left side(P<0.001).Following treatment,all aforementioned indexes decreased significantly,particularly PI on the left side which showed a significant difference from pre-treatment levels(P<0.001).③Before treatment,the levels of serum IL-1β,TNF-α,and CRP in PD college students were significantly higher compared to those in the healthy group.The difference in IL-1β level was statistically significant(P<0.001).After treatment,there was a noticeable decrease in the levels of IL-1β,TNF-α,and CRP.Specifically,IL-1β showed a significant reduction(P<0.01),and this time the comparison with the healthy group did not reveal any significant difference in IL-1β levels.Conclusion The application of warm moxibustion at the Shenque acupoint demonstrates a significant improvement in both the dysmenorrhea pain rating index and severity among college students with primary dysmenorrhea,while also alleviating the severity and duration of associated symptoms.These positive effects may be attributed to warm moxibustion's ability to enhance uterine microcirculation in individuals with primary dysmenorrhea,and ameliorating inflammatory conditions.

Objective:To explore the lived experiences of family members of terminal ICU patients regarding their humanistic care needs and provide theoretical foundations for developing nursing care plans tailored to their needs.Methods:This study was a descriptive qualitative study. From April to December 2023, 16 family members of terminally ill ICU patients in Shanghai East Hospital, Tongji University were selected for semi-structured interviews using purposive sampling method, and the interview data were qualitatively analyzed using Colaizzi 7-step analysis.Results:The humanistic care needs of family members of terminally ill ICU patients can be categorized into five themes, namely, the need to know the condition at the first time; the need to participate in treatment and decision-making; the need to respect the wishes of terminally ill patients; the need for psychological care; and the need for social support.Conclusions:The humanistic care needs of family members of terminal ICU patients remain largely unmet. Nursing professionals should consider these needs and preferences and provide family members with professional guidance to help them establish positive coping mechanisms.

Objective:To systematically review the prediction models for extubation failure in critically ill patients undergoing mechanical ventilation, providing a reference for healthcare professionals in selecting appropriate models to identify high-risk populations.Methods:Literature on the construction of prediction models for extubation failure risk in critically ill patients undergoing mechanical ventilation was retrieved from China National Knowledge Infrastructure, Wanfang Database, VIP, SinoMed, PubMed, Web of Science, Embase, and Cochrane Library. The search was limited from database inception to February 2024. Two researchers independently screened the literature and extracted data, using bias risk assessment tools to evaluate the bias risk and applicability of the prediction models.Results:A total of nine studies were included, with the most common predictive factors being mechanical ventilation duration, Glasgow Coma Scale score, cough reflex strength, age, and 24-hour input/output volume. The area under the receiver operating characteristic curve for the models ranged from 0.689 to 0.926, indicating good predictive performance. However, the risk of bias was high, mainly due to small sample sizes, the selection of predictive factors based on univariate analysis, and lack of proper internal validation.Conclusions:Existing prediction models show good predictive performance, but they carry high bias risk. Future studies should improve research design, adhere to model development and reporting guidelines, and develop well-performing, user-friendly prediction models to more accurately identify high-risk populations for extubation failure.

The 2025 American Society of Clinical Oncology(ASCO)Annual Meeting was held in Chicago from May 30 to June 3,2025.As one of the largest and most influential academic events in global oncology,the ASCO meeting brought together numerous world-class oncology experts.It focused on the unmet clinical needs in gastrointestinal malignancies such as hepatocellular carcinoma,cholangiocarcinoma,and pancreatic cancer,and presented a wealth of cutting-edge research findings and therapeutic innovations.These advances provide important evidence-based support for the diagnosis and treatment of hepatobiliary and pancreatic cancers.Based on the latest achievements presented at ASCO 2025,this article discusses the hot topics and future directions in the management of hepatobiliary and pancreatic tumors.

Objective:To explore the alterations in gray matter morphology and contributing factors in patients with post-infectious olfactory dysfunction (PIOD) using voxel-based morphometry (VBM) and surface-based morphometry (SBM), thereby providing scientific insights into the neuropathological mechanisms underlying PIOD.Methods:A total of 46 PIOD patients (PIOD group) and 46 normosmic volunteers (control group) were recruited from the Smell and Taste Disorders Clinic of Beijing Anzhen Hospital, Capital Medical University, between January 2020 and December 2024. All participants underwent olfactory psychophysical tests (Sniffin′ Sticks) and olfactory event-related potential (oERP) examination. High-resolution T1-weighted 3D MRI structural images were obtained for both groups. VBM was employed to analyze inter-group differences in gray matter volume, while SBM was used to assess cortical thickness and folding index. Correlations between gray matter volume in significant difference brain regions and disease duration, Sniffin′ Sticks scores, oERP parameters were analyzed. A two-tailed P<0.05 was considered statistically significant. Results:No significant differences were observed in age, sex, education level, or Mini-Mental State Examination (MMSE) scores ( t=1.80, χ2=0.41, t=0.17, t=1.77, all P>0.05). Compared with controls, the PIOD group showed significantly lower Sniffin′ Sticks scores ( t=28.70, P<0.001), prolonged oERP latencies and reduced amplitudes (all P<0.001). VBM revealed significantly reduced gray matter volume in the right orbitofrontal cortex (OFC) in the PIOD group ( t=5.38, P<0.001). SBM demonstrated decreased cortical thickness in the right OFC ( t=5.27, P<0.001), with no significant differences in folding index. The gray matter volume in the right OFC was negatively correlated with disease duration ( r=-0.61, P<0.001), but no significant correlations were found with Sniffin′ Sticks scores or oERP parameters. Conclusion:Patients with PIOD show atrophy in the right OFC, which correlates with disease duration, suggesting that persistent olfactory dysfunction may be associated with neurodegenerative changes.

Objective:To systematically review the prediction models for extubation failure in critically ill patients undergoing mechanical ventilation, providing a reference for healthcare professionals in selecting appropriate models to identify high-risk populations.Methods:Literature on the construction of prediction models for extubation failure risk in critically ill patients undergoing mechanical ventilation was retrieved from China National Knowledge Infrastructure, Wanfang Database, VIP, SinoMed, PubMed, Web of Science, Embase, and Cochrane Library. The search was limited from database inception to February 2024. Two researchers independently screened the literature and extracted data, using bias risk assessment tools to evaluate the bias risk and applicability of the prediction models.Results:A total of nine studies were included, with the most common predictive factors being mechanical ventilation duration, Glasgow Coma Scale score, cough reflex strength, age, and 24-hour input/output volume. The area under the receiver operating characteristic curve for the models ranged from 0.689 to 0.926, indicating good predictive performance. However, the risk of bias was high, mainly due to small sample sizes, the selection of predictive factors based on univariate analysis, and lack of proper internal validation.Conclusions:Existing prediction models show good predictive performance, but they carry high bias risk. Future studies should improve research design, adhere to model development and reporting guidelines, and develop well-performing, user-friendly prediction models to more accurately identify high-risk populations for extubation failure.