[摘 要] 目的：探讨异位表达血红蛋白亚基（HBA/HBB）对缺氧条件下嵌合抗原受体T细胞（CAR-T细胞）功能障碍的改善作用及其对肿瘤细胞的杀伤效应。方法：全基因合成技术合成靶向HER2的CAR序列，构建共表达HBA或HBB的CAR慢病毒载体，包装慢病毒后感染人原代T淋巴细胞，制备异位表达HBA/HBB的CAR-T细胞，命名为HBA CAR-T和HBB CAR-T。采用缺氧探针检测小鼠实体瘤缺氧状态。通过流式细胞术检测瘤内CAR-T细胞占比、异位表达血红蛋白亚基的CAR-T细胞阳性率及CAR-T细胞的活性氧、凋亡水平。WB法检测HBA CAR-T和HBB CAR-T内相关血红蛋白亚基表达情况，采用细胞计数板计数检测细胞增殖水平，通过萤光素酶报告基因法检测CAR-T细胞对肿瘤细胞的杀伤能力，qPCR检测CAR-T细胞中缺氧诱导因子-1α（HIF-1α）表达水平，利用MitoXpress Intra试剂盒检测CAR-T细胞内氧气含量。结果：不同细胞构建的实体瘤模型均存在明显缺氧情况，且CAR-T细胞浸润水平与缺氧程度呈显著负相关（P < 0.000 1）。HBA CAR-T与HBB CAR-T构建成功（阳性率 > 60%），相应血红蛋白亚基可稳定表达。缺氧环境下HBA CAR-T和HBB CAR-T的ROS水平、凋亡水平显著下降，增殖、对肿瘤细胞的体外杀伤能力显著强于传统CAR-T细胞（均P < 0.05）。HBA CAR-T与HBB CAR-T内HIF-1α表达降低（均P < 0.001），且缺氧程度显著降低（均P < 0.001）。结论：异位表达血红蛋白亚基可改善缺氧条件下CAR-T细胞功能障碍并增强其对肿瘤细胞的体外杀伤作用。

Objective:To study the diagnostic value and correlation of contrast-enhanced ultrasound time-intensity curve in the main pathological features of cervical cancer.Methods:From August 2021 to August 2023, 100 patients with cervical cancer and 100 patients with benign cervical tumors examined in Dingzhou People’s Hospital were selected for study. Patients with benign cervical tumors were included in the control group, and patients with cervical cancer were included in the observation group.The two groups of patients were examined by PHILIPS EPIQ7 C color Doppler ultrasound diagnostic instrument and PHILIPS EPIQ5 color Doppler ultrasound diagnostic instrument to obtain the quantitative parameters of the time-intensity curve of the two groups. The bivariate Spearman correlation test was used to analyze the main pathological features of cervical cancer patients (lesion diameter, pathological type, vascular invasion, lymph node metastasis, differentiation degree, clinical stage) and the correlation between radiotherapy and chemotherapy and the acquisition of time-intensity curve. The receiver operating characteristic curve (ROC) was drawn to analyze the diagnostic value of the time-intensity curve.Results:The time to peak (TTP) value of the cervical cancer group was lower than that of the control group, and the peak intensity (PI) and maximum peak intensity ratio (PI/PIc) values were higher than those of the control group, the differences were statistically significant ( P<0.05). Compared with patients with non-squamous cell carcinoma, vascular invasion < 1/2, no lymph node metastasis, moderate to high differentiation, stage I-II, and after radiotherapy and chemotherapy, patients with squamous cell carcinoma, vascular invasion ≥ 1/2, lymph node metastasis, low differentiation, stage III-IV, and before radiotherapy and chemotherapy had lower TTP values, higher PI and PI/PIc values, and the differences were statistically significant ( P<0.05). Pathological type, vascular invasion, lymph node metastasis, differentiation degree and clinical stage were negatively correlated with TTP, and positively correlated with PI and PI/PIc ( P<0.05) .Chemoradiotherapy was positively correlated with TTP, and negatively correlated with PI and PI/PIc ( P<0.05). The sensitivity and specificity of combined detection of time intensity curve were higher than those of TTP, PI, PI/Ic, area under curve (AUCTC) and mean transit time (MTT), and the differences were statistically significant ( P<0.05). The AUC values of combined detection of TTP, PI, PI/PIc and time intensity curve were all > 0.85, and the diagnostic value was high. Conclusions:The time-intensity curve of contrast-enhanced ultrasound is closely related to the main pathological features of cervical cancer, such as pathological type, lymph node metastasis and staging. According to the changes, the effect of radiotherapy and chemotherapy can be judged, which can provide a good reference for the diagnosis and treatment of cervical cancer.

OBJECTIVE To study the characteristics,micro-traits and microscopic characteristics of Peucedani radix and seven kinds of its common varieties and summarize the key identification characteristics so as to provide a reference for the effective identifica-tion of Peucedani radix and its common varieties.METHODS The key identification features and high-definition images of Peucedani radix and its common varieties were obtained by using the identification methods of traits,microtraits and microscopy,combined with the techniques of depth-of-field extended imaging and image stitching,and some of the features were digitally extracted and statistical-ly analyzed by SPSS26.0 software.RESULTS The high-definition color image data of Peucedani radix and its common varieties were obtained.Its specific identification features were:root head length and annular sparseness,skin pore shape and area,root texture,and fracture surface oil spot density,etc.under the property identification;the diameter and number of oil chambers,the number of cathe-ters,the presence or absence of bast fibers and wood fibers,etc.under the microscopic identification.The results of statistical analysis showed that there were significant differences in skin pore area,oil chamber diameter and density,and conduit density among different varieties of Peucedani radix(P<0.01).CONCLUSION Micro-traits and microidentification methods can be comprehensively ap-plied to distinguish Peucedani radix and its common varieties.In particular,the microscopic features of polarized light holographic col-or images in cross section have significant distinguishing significance,and some of the features are digitally extracted and statistically analyzed,which makes up for the shortcomings of subjective factors in the traditional empirical identification research,and provides a reference for the circulation,testing,clinical medication,and standard drafting of Peucedani radix.

OBJECTIVE To explore the correlation between phosphatidylinositol-3-kinase alpha(PIK3CA)gene mutation and the clinicopathological characteristics of Epstein-Barr virus(EBV)infection in patients with gastric cancer.METHODS Cancer tissue samples were collected from 593 patients with gastric cancer who under-went surgery at the Affiliated Hospital of Hebei University between Sep.2021 and Sep.2024.EBV infection in pa-tients with gastric cancer was detected by EBV-encoded RNA(EBER)in situ hybridization.Based on the detec-tion results,patients were divided into an EBV-positive group(n=31)and an EBV-negative group(n=562).The incidence of EBV-infected gastric cancer was compared among patients of different genders and ages.The relationship between EBV infection and PIK 3CA gene mutation,as well as clinicopathological character-istics of patients with gastric cancer,was analyzed.Spearman's correlation analysis was used to assess the correla-tion between PIK3CA gene mutation and EBV infection in patients with gastric cancer.RESULTS Among the 593 gastric cancer cases,31 were EBER-positive,with a positive rate of 5.23％.EBER showed scattered dot scope or diffuse staining,appearing as a dark brown signal localized in the nucleus.The PIK3CA gene mutation rate was higher in the EBV-positive group(32.26％)than in the EBV-negative group(P＜0.001).PIK3CA gene muta-tions mainly occurred in exons 9 and 20,with 2 cases of p.E542K mutation,5 cases of p.E545K mutation,1 case of pH1047L mutation,no cases of p.H1047R mutation were detected.The proportion of patients with tumors at the esophagogastric junction,T3-T4 invasion depth,and lymph node metastasis was higher in the EBV-positive group than in the EBV-negative group(P＜0.05).There was a positive correlation between PIK 3CA gene muta-tion and EBV infection in patients with gastric cancer(r=0.742,P=0.026).The proportion of EBV infection was higher in male patients with gastric cancer(6.49％)than in females(P＜0.001).CONCLUSIONS The inci-dence of EBV-infected gastric cancer is low,but it is closely related to PIK3CA gene mutation,gender,tumor lo-cation,invasion depth and lymph node metastasis in patients with gastric cancer.

Objective To establish orthotopic and ectopic pancreatic cancer models in C57BL/6N mice with normal immune function using in vivo imaging technology for visual characterization.Methods Orthotopic and ectopic pancreatic cancer models were established in Kunming mice by injecting a small volume of cell suspension containing firefly luciferase-expressing Panc02-luciferase pancreatic cancer cells into the head of the pancreas or the right axillary region.In vivo imaging technology was used to optimize the modeling method and timing in Kunming mice.Subsequently,the same method was applied to C57BL/6N mice using wild-type Panc02 pancreatic cancer cells to establish orthotopic and ectopic pancreatic cancer models with intact immune function.Key parameters,including body weight,inoculation positive rate,tumor growth time,tumor volume,and pathological characteristics across different organs,were compared be-tween the orthotopic and ectopic models in C57BL/6N mice to evaluate the applicability of these models.Results Both the small animal in vivo imaging experiments in Kunming mice and the tumor growth observation in C57BL/6N mice demonstrated that the construction periods for orthotopic and ectopic pancreatic cancer models were 20 days,with survival rates exceeding 90%.The inoculation positive rates in C57BL/6N mice were 92.3%for the orthotopic model and 78.6%for the ectopic model.On day 20 post-inoculation,the tumor volumes were(117.04±109.56)mm3 for the orthotopic model and(155.68±168.73)mm3 for the ectopic model,indicating high model success rates and consistent tumor growth.HE staining revealed pathological mitotic figures and poorly differentiated tumor tissues in both models of C57BL/6N mice,with no evidence of metastasis to other organs.Conclusions Orthotopic and ectopic pancreatic cancer models in immu-nocompetent mice were successfully developed in this study,mimicking early-stage pancreatic cancer characteristics.These models pro-vide a reliable platform for screening early diagnostic biomarkers and evaluating therapeutic interventions for pancreatic cancer.

OBJECTIVE To establish a closed-loop management system for the whole-process traceability of outpatient drugs based on internet of things(IoT)and blockchain technology,and evaluate its implementation effects.METHODS A closed-loop management system for the whole-process traceability of outpatient drugs covering the entire drug lifecycle was designed using drug traceability codes integrated with IoT and blockchain technology.System effectiveness was evaluated from three dimensions:work efficiency,medication management quality and data safety by comparing indicators such as the acceptance time of incoming drugs and the number of collected drug traceability codes before the system implementation(October to December 2024)and after the system implementation(January to March 2025).RESULTS A closed-loop management system for the whole-process traceability of outpatient drugs,centered around the drug traceability code management system,was successfully established.The acceptance time for incoming drugs was shortened from(4.65±0.26)h before implementation to(0.34±0.08)h after implementation(P＜0.05).The number of collected drug traceability codes increased from 419 018 to 1 236 522,and the coverage rate of traceability codes rose from 28.36%to 89.88%(P＜0.05).The time pharmacists spent on drug expiry management per week decreased from(128.40±19.20)min to(0.56±0.13)min(P＜0.05),and the dispensing time for a single prescription(excluding a part of injections and repackaged drugs)was reduced from(143.25±17.67)s to(15.24±10.08)s(P＜0.05).The time for drug return was reduced from 129.90(122.32,137.00)s to 104.36(89.91,117.33)s(P＜0.05);the number of drug dispensing errors decreased from 2 cases to 0 cases.After the system was launched,there were no data security incidents in our outpatient pharmacy.CONCLUSIONS The constructed closed-loop management system for the whole-process traceability of outpatient drugs can significantly enhance drug traceability accuracy and drug management quality,improve pharmacist work efficiency,and reduce drug management risks,thus providing a feasible solution for the digital transformation of hospital pharmaceutical services.

BACKGROUND: Arsenic trioxide (ATO) is indicated as a broad-spectrum medicine for a variety of diseases, including cancer and cardiac disease. While the role of ATO in hepatic ischemia/reperfusion injury (HIRI) has not been reported. Thus, the purpose of this study was to identify the effects of ATO on HIRI. METHODS: In the present study, we established a 70% hepatic warm I/R injury and partial hepatectomy (30% resection) animal models in vivo and hepatocytes anoxia/reoxygenation (A/R) models in vitro with ATO pretreatment and further assessed liver function by histopathologic changes, enzyme-linked immunosorbent assay, cell counting kit-8, and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay. Small interfering RNA (siRNA) for extracellular signal-regulated kinase (ERK) 1/2 was transfected to evaluate the role of ERK1/2 pathway during HIRI, followed by ATO pretreatment. The dynamic process of autophagic flux and numbers of autophagosomes were detected by green fluorescent protein-monomeric red fluorescent protein-LC3 (GFP-mRFP-LC3) staining and transmission electron microscopy. RESULTS: A low dose of ATO (0.75 μmol/L in vitro and 1 mg/kg in vivo ) significantly reduced tissue necrosis, inflammatory infiltration, and hepatocyte apoptosis during the process of hepatic I/R. Meanwhile, ATO obviously promoted the ability of cell proliferation and liver regeneration. Mechanistically, in vitro  studies have shown that nontoxic concentrations of ATO can activate both ERK and phosphoinositide 3-kinase-serine/threonine kinase (PI3K-AKT) pathways and further induce autophagy. The hepatoprotective mechanism of ATO, at least in part, relies on the effects of ATO on the activation of autophagy, which is ERK-dependent. CONCLUSION Low, non-toxic doses of ATO can activate ERK/PI3K-AKT pathways and induce ERK-dependent autophagy in hepatocytes, protecting liver against I/R injury and accelerating hepatocyte regeneration after partial hepatectomy.
Animals ; Arsenic Trioxide ; Autophagy/physiology* ; Reperfusion Injury/prevention & control* ; Mice ; Male ; Proto-Oncogene Proteins c-akt/physiology* ; Arsenicals/therapeutic use* ; Oxides/therapeutic use* ; Liver/metabolism* ; Extracellular Signal-Regulated MAP Kinases/metabolism* ; Mice, Inbred C57BL

ObjectiveTo explore the clinical safety of Feilike Mixture （FLK） in the real world. MethodsThe safety of all children who received FLK from 29 institutions in 12 provinces between January 21，2021 and December 25，2021 was evaluated through prospective centralized surveillance and a nested case control study. ResultsA total of 3 035 juveniles were included. There were 29 research centers involved，which are distributed across 12 provinces，including one traditional Chinese medicine （TCM） hospital and 28 general hospitals. The average age among the juveniles was （4.77±3.56） years old，and the average weight was （21.81±12.97） kg. Among them，119 cases （3.92%） of juveniles had a history of allergies. Acute bronchitis was the main diagnosis for juveniles，with 1 656 cases （54.46%）. FLK was first used in 2 016 cases （66.43%），and 142 juvenile patients had special dosages，accounting for 4.68%. Among them，92 adverse drug reactions （ADRs） occurred，including 73 cases of gastrointestinal system disorders，10 cases of metabolic and nutritional disorders，eight cases of skin and subcutaneous tissue diseases，two cases of vascular and lymphatic disorders，and one case of systemic diseases and various reactions at the administration site. The manifestations of ADRs were mainly diarrhea，stool discoloration，and vomiting，and no serious ADRs occurred. The results of multi-factor analysis indicated that special dosages （the use of FLK）［odds ratio （OR） of 2.642， 95% confidence interval （CI） of 1.105-6.323］，combined administration： spleen aminopeptide （OR of 4.978， 95%CI of 1.200-20.655），and reason for combined administration： anti-infection （OR of 1.814， 95%CI of 1.071-3.075） were the risk factors for ADRs caused by FLK. Conclusion92 ADRs occurred among 3 035 juveniles using FLK. The incidence of ADRs caused by FLK was 3.03%，and the severity was mainly mild or moderate. Generally，the prognosis was favorable after symptomatic treatment such as drug withdrawal or dosage reduction，suggesting that FLK has good clinical safety.

Bisphenol A(BPA)is an environmental endocrine disruptor commonly found in industri-al products such as plastics,resin coatings,and paper coatings,and it poses potential reproductive hazards.Despite extensive research,studies examining its effects on the F1 generation of rabbits are limited.This study established a BPA exposure model in pregnant female rabbits to investigate its impact on reproductive hormones,apoptosis,oxidative stress,inflammatory responses,and tissue integrity in weaned female offspring.The results indicate that BPA exposure induces oxidative stress and inflammation in F1 rabbits,disrupts hormonal balance,and affects antioxidant enzymes and inflammatory mediators through the modulation of the Nrf2 and NF-κB signaling pathways,ultimately leading to apoptosis and tissue damage.

Reactive oxygen species (ROS) are major contributors to radiation therapy-induced side effects in cancer patients. A fusion antioxidant enzyme comprising glutathione S-transferase (GST), superoxide dismutase 1 (SOD1), and a transmembrane peptide has been shown to effectively mitigate ROS-induced damage. To enhance its targeting capability, the fusion protein was further modified by incorporating a matrix metalloproteinase-2/9 substrate peptide (X) and the transmembrane peptide R9, yielding the antioxidant enzyme GST-SOD1-X-R9 (GS1XR). This modification reduced its transmembrane ability in tumor cells, thereby selectively protecting normal cells from oxidative stress. However, the use of non-human GST poses potential immunogenicity risks. In this study, we employed seamless cloning technology to construct an expression vector containing the human GST gene to replace the non-human GST gene, and then expressed and purified novel fusion antioxidant enzymes GS1R and GS1XR. The protective effects of newly constructed GS1R and GS1XR against hydrogen peroxide (H₂O₂)-induced oxidative damage in L-02 cells were then evaluated using GS1 as a control. Enzymatic activity assays revealed that the specific activity of GST in GS1XR remained unchanged compared to the unmodified protein, while SOD activity was enhanced. Exposure to 200 μmol/L H₂O₂ transiently activated the nuclear factor-erythroid 2-related factor 2 (Nrf2) pathway; however, this activation diminished after 24 h, reducing cell viability to 48.4%. Both GS1R and GS1XR effectively scavenged intracellular ROS, directly counteracting oxidative stress and promoting Nrf2 nuclear translocation, thereby activating antioxidant pathways and restoring cell viability to normal levels. The two enzymes showed comparable efficacy. In contrast, GS1, lacking transmembrane capability, was restricted to scavenging extracellular ROS and provided only limited protection. In conclusion, both novel fusion antioxidant enzymes demonstrated significant potential in safeguarding normal cells from ROS-mediated oxidative damage. The findings provide a foundation for further investigation in related field.
Humans ; Oxidative Stress/drug effects* ; Hydrogen Peroxide ; Antioxidants/metabolism* ; Glutathione Transferase/metabolism* ; Recombinant Fusion Proteins/pharmacology* ; Superoxide Dismutase-1 ; Reactive Oxygen Species/metabolism* ; Superoxide Dismutase/biosynthesis*