[摘 要] 目的：探讨乙醛脱氢酶1A1（ALDH1A1）mRNA负载树突状细胞（DC）疫苗体外诱导抗鼻咽癌干细胞免疫反应的能力。方法：体外转录ALDH1A1 mRNA并电转染至人外周血DC，构建ALDH1A1-DC疫苗，转染EGFP mRNA为对照。流式细胞术检测DC表型、淋巴细胞亚群、细胞毒性T 淋巴细胞（CTL）增殖和细胞因子分泌。分选ALDH+和ALDH- C666-1细胞，检测干细胞标志物（CD24、CD133）和干性基因（OCT4、SOX2、Nanog）的表达水平，采用CCK-8法评估ALDH1A1-DC疫苗诱导的特异性免疫杀伤能力。结果：EGFP-DC荧光表达率显著高于未转染DC（P < 0.000 1），ALDH1A1-DC抗原提呈分子HLA-ABC、共刺激分子CD86、CD40和CCR7表达与未转染DC相比显著增强（P = 0.001 3、P = 0.005 9、P = 0.000 4及P = 0.001 9）。与单独淋巴细胞组（T）及空载DC诱导的淋巴细胞组（DC-T）相比，ALDH1A1-DC诱导的CTL比例最高（均P < 0.000 1），分裂指数最高（P = 0.000 2、P = 0.000 3），且CD8+CD69+（P = 0.000 5、P < 0.000 1）和CD8+ IFN-γ+（P = 0.012 6、P = 0.001 9）细胞亚群比例均显著提高。ALDH1A1-DC-T细胞对ALDH+C666-1悬浮细胞球的杀伤效果显著优于ALDH-细胞球（P = 0.001 0），且与其他各组淋巴细胞相比表现出较强的抗ALDH+C666-1悬浮细胞球的免疫效应（P = 0.001 0，P < 0.000 1）。然而，HLA-ABC阻断后其对ALDH+C666-1细胞的特异性杀伤效应显著降低（P < 0.000 1）。结论：mRNA转染DC后可翻译蛋白并促进DC成熟，增强其诱导特异性CTL免疫反应的能力。

Objective:This study aimed to analyze the infection status of viral viruria within one year after kidney transplantation, its impact on renal allograft function, and associated risk factors.Methods:A retrospective case-control study was conducted, involving 370 patients who underwent allogeneic kidney transplantation at Renji Hospital, Shanghai Jiao Tong University School of Medicine, from January 1, 2020 to December 31, 2021. Urinary viral loads of BK virus (BKV), JC virus (JCV), cytomegalovirus (CMV), and Epstein-Barr virus (EBV) were detected using PCR fluorescent probe assays. Patients were categorized into infection and non-infection groups. Glomerular filtration rate (GFR) and tacrolimus trough concentration was measured during infections, and clinical data were collected. Univariate analysis was performed to identify risk factors for viral viruria.Results:The 1-year patient survival rate and graft survival rate were both 98.6%. The incidence rates of viral viruria were as follows: JCV (42.7%), BKV (29.7%), CMV (11.6%), and EBV (2.9%), with statistically significant differences among viruses ( P<0.001). Single viral infection accounted for 48% of cases, while co-infections were predominantly BKV+JCV (9%). JCV infection rates remained consistently high throughout the year (22.4%-28.9%), whereas BKV infections peaked at 3 months postoperatively (20.5%). Co-infection with low-load JCV (>2 000 copies/ml) and CMV (>6 000 copies/ml) led to a significant decline in GFR at 6 months post-transplantation [median difference: 16.7 ml/(min×1.73 m2), P=0.019]. Univariate analysis revealed that elevated tacrolimus trough concentration was independent risk factor for BKV (4.90 vs. 4.30 ng/ml, Z=4.29, P<0.001) and JCV infections (5.30 vs. 4.80 ng/ml, Z=4.25, P<0.001). Conclusion:High incidences of JCV and BKV infections were observed post-kidney transplantation. Co-infection with low-load JCV and CMV accelerates renal function impairment, highlighting the critical role of tacrolimus concentration management in reducing viral infection risks.

Objective:To establish a serum detection method of surface-enhanced Raman spectroscopy (SERS) combining with machine learning for early screening of colorectal cancer (CRC).Methods:Serum samples were collected from 150 CRC patients diagnosed at Jiangdu People′s Hospital, Affiliated to Yangzhou University, and also from 37 healthy subjects. Gold nanohexapod (AuNHs) arrays were prepared using an oil-water interface self-assembly method. A 5 μl serum sample was applied onto the AuNHs array. Scatheless and rapid detection for serum were performed using a Renishaw inVia Raman spectrometer at room temperature with a laser wavelength of 785 nm, exposure time of 10 s, and power of 5 mW. The raw SERS spectra were preprocessed using Savitzky-Golay smoothing, AsLS baseline correction, and Min-Max normalization with Origin 2019 software. Furthermore, the principal component analysis (PCA)-support vector machine (SVM) model was constructed using Python′s scikit-learn library. Leave-One-Out Cross-Validation (LOOCV) was used to evaluate the model′s accuracy, sensitivity, specificity, and area under the curve (AUC).Results:The AuNHs arrays exhibited uniform morphology. The relative standard deviation (RSD) of the SERS intensity at 1 080 cm -1 was 5.69%, and the RSD of the SERS intensity at 1 340 cm -1 was 6.20%. The limit of detection (LOD) of the AuNHs array was 9.42×10 -12 mol/L. The PCA-SVM model achieved an accuracy of 90.91% (170/187), sensitivity of 96.79% (181/187), specificity of 99.47% (186/187), and an AUC of 0.98. The most significant characteristic peaks distinguishing different CRC stages were at 747, 940, 1 000, 1 447, and 1 612 cm -1. Conclusion:The serum detection method based on SERS combined with machine learning can accurately screen CRC with higher accuracy, sensitivity, and specificity, demonstrating potential clinical application value.

Pelvic organ prolapse (POP), whose etiology is influenced by genetic and clinical risk factors, considerably impacts women's quality of life. However, the genetic underpinnings in non-European populations and comprehensive risk models integrating genetic and clinical factors remain underexplored. This study constructed the first polygenic risk score (PRS) for POP in the Chinese population by utilizing 20 disease-associated variants from the largest existing genome-wide association study. We analyzed a discovery cohort of 576 cases and 623 controls and a validation cohort of 264 cases and 200 controls. Results showed that the case group exhibited a significantly higher PRS than the control group. Moreover, the odds ratio of the top 10% risk group was 2.6 times higher than that of the bottom 10%. A high PRS was significantly correlated with POP occurrence in women older than 50 years old and in those with one or no childbirths. As far as we know, the integrated prediction model, which combined PRS and clinical risk factors, demonstrated better predictive accuracy than other existing PRS models. This combined risk assessment model serves as a robust tool for POP risk prediction and stratification, thereby offering insights into individualized preventive measures and treatment strategies in future clinical practice.
Humans ; Female ; Pelvic Organ Prolapse/epidemiology* ; Middle Aged ; Risk Assessment/methods* ; China/epidemiology* ; Multifactorial Inheritance ; Aged ; Risk Factors ; Genome-Wide Association Study ; Genetic Predisposition to Disease ; Case-Control Studies ; Adult ; Polymorphism, Single Nucleotide ; Genetic Risk Score ; East Asian People

This study identified six novel azaphilones, isochromophilones G-L (1-6), and three novel biosynthetically related congeners (7-9) from Diaporthe sp. SCSIO 41011. The structures and absolute configurations were elucidated through comprehensive spectroscopic analyses combined with experimental and calculated electronic circular dichroism (ECD) spectra. Significantly, three highly oxygenated azaphilones contain an acetyl group at the terminal chain (4) or linear conjugated polyenoid moieties (5 and 6), which occur infrequently in the azaphilone family. Additionally, several compounds demonstrated inhibition of lipopolysaccharide (LPS)-induced nuclear factor kappa-B (NF-κB) activation in RAW 264.7 macrophages at 20 μmol·L^-1. The novel compound (1) effectively inhibited receptor activator of NF-κB ligand (RANKL)-induced osteoclast differentiation without exhibiting cytotoxicity in bone marrow and RAW 264.7 macrophages, indicating its potential as a promising lead compound for osteolytic disease treatment. This research presents the first documented evidence of azaphilone derivatives as inhibitors of RANKL-induced osteoclastogenesis.
Animals ; Mice ; RANK Ligand/genetics* ; RAW 264.7 Cells ; Osteoclasts/metabolism* ; Benzopyrans/isolation & purification* ; Osteogenesis/drug effects* ; Macrophages/metabolism* ; Molecular Structure ; Pigments, Biological/isolation & purification* ; Ascomycota/chemistry* ; NF-kappa B/genetics* ; Cell Differentiation/drug effects*

Objective:To discuss the ameliorative effect of total flavonoids from corn silk(TFCS)on kidney injury in the rats with urate nephropathy,and to clarify the possible mechanism.Methods:Sixty male Wistar rats were randomly divided into control group,model group,positive control group[benzbromarone(BZM)group,5 mg·kg-1·d-1],low dose of TFCS group(20 mg·kg-1·d-1),medium dose of TFCS group(40 mg·kg-1·d-1),and high dose of TFCS group(80 mg·kg-1·d-1),and there were 10 rats in each group.Except for control group,the rats in the other groups were administered potassium oxonate(350 mg·kg-1)and adenine(70 mg·kg-1)by gavage for 4 weeks to establish the hyperuricemic nephropthy models.The rats in different doses of TFCS groups were treated with TFCS for 2 weeks.Speckle blood flow imager was used to detect the renal blood perfusion of the rats in various groups and the kidney coefficients of the rats in various groups were caculated;HE staining and Masson staining were used to observe the pathomorphology and fibrosis degrees of kidney tissue of the rats in various groups and enzyme-linked immunosorbent assay(ELISA)method was used to detect the levels of uric acid(UA),creatinine(Cr),blood urea nitrogen(BUN),interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α)in the serum and levels of β2-microglobulin(β2-MG)and microalbumin(ALB)in the urine of the rats in various groups;Western blotting method was used to detect the expression levels of urate transporter 1(URAT1),glucose transporter 9(GLUT9),and ATP-binding cassette transporter G2(ABCG2)proteins in kidney tissue of the rats in various groups.Results:Compared with control group,the renal blood perfusion volume of the rats in model group was significantly decreased(P<0.01).Compared with model group,the renal blood perfusion volumes of the rats in BZM group and low,medium,and high doses of TFCS groups were significantly increased(P<0.05 or P<0.01).Compared with control group,the kidney weight of the rats in model group was increased,with visible white granular spots on the surface,absence of blood color,and kidney volume was increased.Compared with model group,the kidney volumes of the rats in BZM group and medium and high doses of TFCS groups were decreased,with color tending toward that in control group,and the white granular spots on the surface were significantly reduced.Compared with model group,the kidney coefficients of the rats in BZM group and medium and high doses of TFCS groups were decreased(P<0.01).The HE staining results showed there were no abnormalities in kidney tissue structure in control group,while there were a small amount of brown-yellow urate crystal deposition and interstitial connective tissue hyperplasia in model group;compared with model group,the kidney tissue damage and inflammatory infiltration were alleviated to varying degrees in BZM group and different doses of TFCS groups.The Masson staining results revealed no obvious collagen fiber deposition in control group,whereas significant blue collagen fiber deposition in kidney tissue of the rats was found in model group,and the collagen volume fraction(CVF)was increased compared with control group(P<0.01);compared with model group,the CVFs of the rats in BZM group and different doses of TFCS groups were decreased(P<0.01).The ELISA results showed that compared with control group,the levels of UA,Cr,BUN,IL-6,and TNF-α in serum of the rats in model group were increased(P<0.01);compared with model group,the levels of UA,Cr,BUN,IL-6,and TNF-α in serum of the rats in BZM group and medium and high doses of TFCS groups were decreased(P<0.01).Compared with control group,the levels of β2-MG and ALB in urinary in model group were increased(P<0.01);compared with model group,the levels of β2-MG and ALB in urinary of the rats in different doses of TFCS groups were decreased(P<0.05 or P<0.01).The Western blotting results showed that compared with control group,the expression levels of URAT1 and GLUT9 proteins in kidney tissue of the rats in BZM group and model group were increased(P<0.01),while the expression level of ABCG2 protein was decreased(P<0.01).Compared with model group,the expression levels of URAT1 and GLUT9 proteins in kidney tissue of the rats in different doses of TFCS groups were decreased(P<0.05 or P<0.01),and the expression level of ABCG2 protein was increased(P<0.01).Conclusion:TFCS can significantly alleviate the kidney injury in the rats with urate nephropathy model,and its mechanism may be related to the downregulation of expression of URAT1 and GLUT9 proteins and upregulation of ABCG2 protein expression in kidney tissue.

Objective To investigate the role and potential molecular mechanisms of insulin-like growth factor 2 mRNA-binding protein 3(IGF2BP3)in preeclampsia(PE).Methods The expres-sion of IGF2BP3 in placental tissues from patients with PE was detected using quantitative reverse transcription polymerase chain reaction.IGF2BP3 was knocked down via RNA interference technology to evaluate its effects on the proliferation,invasion,apoptosis and cell cycle of HTR-8/SVneo and JEG-3 trophoblast cell lines.Transcriptome sequencing was employed to analyze changes in cellular biological functions and the stability of the potential downstream molecule circPAPPA following IGF2BP3 interference.RNA binding protein immunoprecipitation(RIP)and fluorescence in situ hy-bridization(FISH)were utilized to validate the direct targeting relationship between IGF2BP3 and circPAPPA.Results IGF2BP3 was significantly downregulated in PE placentas.Knockdown of IGF2BP3 inhibited trophoblast cell proliferation and invasion,promoted cell apoptosis and cellcycle arrest.Functional enrichment analysis revealed that IGF2BP3 was involved in invasion and hypoxia response,regulating signaling pathways such as phosphatidylinositol-3-kinase(PI3K),mitogen-activated protein kinase(MAPK)and hypoxia-inducible factor-1(HIF-1).Knockdown of IGF2BP3 led to a reduction in circPAPPA stability,and RIP and FISH experiments confirmed the direct targeting rela-tionship between IGF2BP3 and circPAPPA.Conclusion IGF2BP3 is downregulated in PE and reg-ulates the stability of circPAPPA in an N6-adenosine methylation(m6A)-dependent manner,there-by affecting trophoblast cell function.

[Objective]To explore the effect and potential mechanism of quercetin on polarization of lipopolysaccharide(LPS)-induced primary microglia.[Methods]Primary rat microglia were isolated and cultured,and then randomly divided into control group,model group and quercetin low-dose,medium-dose,high-dose groups.In model group,microglial activation was induced with LPS.In the quercetin groups,microglia were pretreated with quercetin at concentrations of 20,40 and 80 μmol·L-1 for 1 hour,followed by the addition of LPS to the culture medium for an additional 24 hours.Cell viability was assessed by using the CCK-8 assay.The nitric oxide(NO)content in the supernatant was measured by the Griess assay.Microglia activation was detected by ionized calcium binding adapter molecule 1(Iba1)/CD68 immunofluorescence staining.The expression levels of CD86,inducible nitric oxide synthase(iNOS),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),IL-6,CD206,arginase-1(Arg-1),chitinase like protein 1/2(Ym1/2),IL-10 and transforming growth factor-β(TGF-β)mRNA were determined by Real time-quantitative polymerase chain reaction(RT-qPCR).Network pharmacology and molecular docking methods were employed to predict the potential mechanisms of quercetin in regulating microglia polarization.The protein expression levels of CD86,iNOS,CD206,Arg-1,phosphophorylated-phosphatidylinositol 3-kinase(p-PI3K)/phosphatidylinositol 3-kinase(PI3K),phosphophorylated-protein kinase B(p-Akt)/protein kinase B(Akt),phosphophorylated-nuclear factor-κB(p-NF-κB)/nuclear factor-κB(NF-κB)and phosphophorylated-inhibitor of NF-κB α(p-IκB-α)/inhibitor of NF-κB α(IκB-α)were determined by Western blot.[Results]Compared with LPS group,the NO release and CD68 mean fluorescence intensity of microglia in quercetin groups were significantly reduced(P<0.01),indicating that quercetin inhibited LPS-induced activation of primary microglia.Quercetin inhibited the mRNA and protein expression of CD86 and iNOS(P<0.05,P<0.01),and decreased the mRNA expression of pro-inflammatory factors TNF-α,IL-1β and IL-6(P<0.05,P<0.01).Additionally,quercetin promoted the mRNA and protein expression of CD206 and Arg-1(P<0.05,P<0.01),and upregulated the mRNA expression of anti-inflammatory factors Ym1/2,IL-10 and TGF-β(P<0.05,P<0.01).These findings indicated that quercetin promoted the transformation of LPS-induced primary microglia from the M1 type to the M2 type,thereby inhibiting neuroinflammation.The results of network pharmacology and molecular docking indicated that the regulation of microglia polarization by quercetin may be through the PI3K/Akt/NF-κB signaling pathway.In vitro experimental results showed that compared with LPS group,the protein expression levels of p-NF-κB and p-IκB-α in quercetin medium and high dose groups were significantly reduced(P<0.01),and the protein expression of p-PI3K and p-Akt was significantly increased(P<0.05,P<0.01).[Conclusion]Quercetin inhibited LPS-induced microglia inflammatory response and promoted microglia polarization to the M2 type,and the mechanism may be related to its regulation of the PI3K/Akt/NF-κB signaling pathway.

Objective:To explore the prognostic differences between liver resection and transarterial chemoembolization (TACE) in patients with Budd-Chiari syndrome (BCS) complicated by hepatocellular carcinoma (HCC) and to identify independent risk factors affecting patient survival.Methods:The clinical and follow-up data of 103 patients with stage Ⅰa-Ⅲa BCS complicated by HCC treated at the First Affiliated Hospital of Zhengzhou University from Aug 2015 to Sep 2023 were retrospectively analyzed.Results:Patients were divided into two groups based on their initial treatment choices: the liver resection group ( n=20) and the TACE group ( n=83). Before propensity score matching(PSM), the median overall survival in the liver resection group was 42 months longer than in the TACE group (74 months vs. 32 months, P=0.002). After PSM, the median overall survival remained significantly longer in the liver resection group by 39 months (74 months vs. 35 months, P=0.032). In terms of disease-free survival, before PSM, the liver resection group was 30-month longer than the TACE group (42 months vs. 12 months, P=0.001). After PSM, the difference in median disease-free survival between the two groups was 23 months (35 months vs. 12 months, P=0.018). Multivariate Cox regression analysis identified treatment modality and maximum tumor diameter as independent risk factors for overall survival, while treatment modality was the only independent factor for disease-free survival. Conclusions:Liver resection significantly prolongs both overall survival and disease-free survival in resectable HCC in BCS patients compared to TACE. Treatment modality and tumor size are key prognostic factors influencing overall survival.

Objective:To evaluate the feasibility of training physicians from secondary comprehensive hospitals in the clinical assessment of brain death and to provide recommendations for nationwide implementation.Methods:This prospective cohort study enrolled physicians who completed standardized training in clinical brain death determination at five pilot hospitals between June and December 2023. Participants were from internal medicine, neurology, critical care, emergency, or anesthesiology departments of secondary comprehensive hospitals and had ≥5 years of clinical experience. Organ donation coordinators and surgeons involved in organ donation or transplantation were excluded. The training program comprised four modules: didactic lectures, bedside demonstrations, simulation-based practice, and written theoretical assessment with review. The theoretical assessment was considered qualified if the score was 60 or above. Participants were categorized into ≥80 and <80 groups based on assessment scores. Between-group comparisons were conducted using rank-sum or chi-square tests.Results:A total of 191 physicians from 74 secondary comprehensive hospitals were enrolled. Most held a bachelor's degree [89.5%(171/191)] and had intermediate [47.1%(90/191)] or associate senior [36.1%(69/191)] professional titles; [59.7%(114/191)] were from non-neurology specialties. The overall pass rate was 99.5% (190/191), with a mean score of 82.4±7.1. Compared with those scoring<80 (56 participants), physicians scoring ≥80 (135 participants) differed significantly by professional title, province, and department ( P=0.014, 0.019 and 0.039). The proportion scoring<80 was higher among junior/intermediate versus senior titles [38.0%(41/108) vs 18.1%(15/83), P=0.003), and among non-neurology/critical care departments (emergency, internal medicine, anesthesiology) versus neurology/critical care [39.7%(31/78) vs 22.1%(25/113), P=0.009]. Only 2.09%(4/191) achieved a perfect score. Across all test items, the overall error rate was 14.99%(700/4 670). The five knowledge points with the highest error rates were mistriggering of mechanical ventilation [96.97%(32/33)], corneal reflex [42.25%(30/71)], spinal reflexes [24.25%(65/268)], documentation of the determination [21.21%(7/33)], and the apnea test procedure [20.73%(57/275)]. Conclusions:The pilot hospitals can effectively deliver clinical training for brain death determination, supporting nationwide promotion. However, physicians' theoretical grounding in neurology at secondary comprehensive hospitals appears relatively weak. Training curricula should be optimized to further improve training quality.