Objective To investigate whether fenoldopam(FNDP)(an agonist of type 1 dopamine receptor)has a protective effect on thoracic aortic aneurysm(TAA)in mice.Methods Three-week-old male C57BL/6J mice were treated with β-aminopropionitrile(BAPN)to induce TAA.The mice were divided into three groups:the con-trol group,the BAPN group,and the BAPN+FNDP group(FNDP injected intraperitoneally).The incidence and survival rate of TAA were recorded.Gross anatomy of the whole aortae was observed.Elastin staining was per-formed to assess morphological change,while immunohistochemistry was employed to evaluate the expressions of matrix metalloproteinase 2(MMP2),matrix metalloproteinase 9(MMP9)and cluster of differentiation 68(CD68)respectively.Gelatin zymography was conducted to assess MMP2 and MMP9 activity.Reverse transcription-poly-merase chain reaction(RT-PCR)was performed to measure the mRNA expression levels of dopamine receptor D1(D1DR),dopamine receptor d2(D2DR),dopamine receptor d3(D3DR),dopamine receptor d5(D5DR),in-terleukin-1β(IL-1β),interleukin-6(IL-6),tumour necrosis factor-α(TNF-α),monocyte chemoattractant pro-tein-1(MCP-1),alpha-smooth muscle actin(α-SMA)and smooth muscle protein 22-alpha(SM22α).Results Compared to the control group,the BAPN group exhibited significant formation of TAA.Elastic fiber disruption was also observed in the thoracic aortic wall,along with a significant decrease in the mRNA levels of D1DR and D5DR.The BAPN+FNDP group showed a significant reduction in the incidence of TAA formation and the rate of aneu-rysm rupture compared to the BAPN group.The disruption and rupture of elastic fibers in the thoracic aortic wall were significantly improved in the BAPN+FNDP group.The levels of MMP2 and MMP9 in the thoracic aortic wall significantly decreased,and the enzymatic activity of MMP2 in the serum was significantly reduced.Moreover,macrophage infiltration in the thoracic aortic wall was significantly reduced and the mRNA levels of IL-1β,IL-6,TNF-α and MCP-1 also significantly decreased after FNDP treatment.There was no statistically significant differ-ence in the mRNA levels of α-SMA and SM22α.Conclusion FNDP shows an inhibitory effect on TAA progres-sion in mice,suggesting a potential of FNDP as a therapeutic agent for TAA.

A retrospective analysis was conducted on a MonoMAC syndrome case admitted in October 2022 to the First Affiliated Hospital of Zhejiang University School of Medicine. The patient, a 16-year-old female with a history of persistent monocytopenia and mild anemia for several years, experienced recurrent symptoms of cough, expectoration, and fever, leading to multiple visits to the hospital. The diagnosis of MonoMAC syndrome was confirmed through comprehensive assessments including routine blood tests, pathogen metagenomic sequencing, lung and bone marrow biopsies, and next-generation sequencing of peripheral blood. The patient underwent haploidentical hematopoietic stem cell transplantation, with a smooth course of transplantation, achieving neutrophil engraftment on + 16 d and platelet engraftment on + 17 d, eventually restoring normal monocyte and NK cell counts. MonoMAC syndrome patients often initially present with infectious symptoms, and the diagnosis can be established based on significant monocytopenia in routine blood tests, history of non-tuberculous mycobacterial infections, and GATA2 germline mutations. Allogeneic hematopoietic stem cell transplantation may be required for some patients to improve their prognosis.

Objective:To investigate the value of systemic inflammatory response syndrome (SIRS), pediatric sequential organ failure assessment (pSOFA) and pediatric critical illness score (PCIS) in predicting mortality of pediatric sepsis in pediatric intensive care units (PICU) from Southwest China.Methods:This was a prospective multicenter observational study. A total of 447 children with sepsis admitted to 12 PICU in Southwest China from April 2022 to March 2023 were enrolled. Based on the prognosis, the patients were divided into survival group and non-survival group. The physiological parameters of SIRS, pSOFA and PCIS were recorded and scored within 24 h after PICU admission. The general clinical data and some laboratory results were recorded. The area under the curve (AUC) of the receiver operating characteristic curve was used to compare the predictive value of SIRS, pSOFA and PCIS in mortality of pediatric sepsis.Results:Amongst 447 children with sepsis, 260 patients were male and 187 patients were female, aged 2.5 (0.8, 7.0) years, 405 patients were in the survival group and 42 patients were in the non-survival group. 418 patients (93.5%) met the criteria of SIRS, and 440 patients (98.4%) met the criteria of pSOFA≥2. There was no significant difference in the number of items meeting the SIRS criteria between the survival group and the non-survival group (3(2, 4) vs. 3(3, 4) points, Z=1.30, P=0.192). The pSOFA score of the non-survival group was significantly higher than that of the survival group (9(6, 12) vs. 4(3, 7) points, Z=6.56, P<0.001), and the PCIS score was significantly lower than that of the survival group (72(68, 81) vs. 82(76, 88) points, Z=5.90, P<0.001). The predictive value of pSOFA (AUC=0.82) and PCIS (AUC=0.78) for sepsis mortality was significantly higher than that of SIRS (AUC=0.56) ( Z=6.59, 4.23, both P<0.001). There was no significant difference between pSOFA and PCIS ( Z=1.35, P=0.176). Platelet count, procalcitonin, lactic acid, albumin, creatinine, total bilirubin, activated partial thromboplastin time, prothrombin time and international normalized ratio were all able to predict mortality of sepsis to a certain degree (AUC=0.64, 0.68, 0.80, 0.64, 0.68, 0.60, 0.77, 0.75, 0.76, all P<0.05). Conclusion:Compared with SIRS, both pSOFA and PCIS had better predictive value in the mortality of pediatric sepsis in PICU.

Objective To explore the application effect of air bag compression combined with multi-step automatic decompression in radial artery hemostasis after coronary angiography in elderly patients. Methods A total of 630 elderly patients who underwent transradial coronary angiography in our hospital from March 2023 to April 2024 were selected as study subjects and randomly divided into control group, observation group 1, and observation group 2, with 210 patients in each group. The control group received manual compression hemostasis with a three-step air bag compressor after surgery, while the observation group 1 and observation group 2 received automatic compression hemostasis with a customized pressure step compressor for three-step and five-step compression, respectively, on the air bag compressor after surgery. The bleeding rate, delayed pressure release rate, radial artery stenosis rate, radial artery occlusion rate, and nursing work time were compared among the three groups after surgery (after CAG and before removal of the air bag compressor). Pain scores, heart rate, and blood pressure were recorded after surgery and before pressure release at 1 h after surgery. Results The observation group 2 had better results in bleeding rates, delayed pressure release rates, radial artery stenosis rates, and occlusion rates at the puncture site compared with the control group and observation group 1(P < 0.05). The nursing work time in the observation group 1 and observation group 2 was shorter than that in the control group (P < 0.05). The pain scores, heart rate, and systolic blood pressures before pressure release at 1 h after surgery were better in all three groups compared with those after surgery (P < 0.05). The pain scores, heart rates, and blood pressures before pressure release at 1 h after surgery in the observation group 2 were better than those in the control group and observation group 1 (P < 0.05). Conclusion The application of air bag compression combined with a five-step gradual automatic decompression method for compression hemostasis can effectively reduce the incidence of radial artery complications, shorten nursing work time, and improve patients' comfort.

Objective To investigate the impact of selective ulnar artery continuous circulatory compression in reducing radial artery occlusion in elderly patients undergoing coronary intervention for coronary artery disease. Methods A total of 450 elderly patients with coronary heart disease undergoing coronary intervention were selected as study objects, and were randomly divided into control group and experimental group, with 225 patients in each group. The control group received non-occlusive radial artery compression postoperatively, while the experimental group underwent selective ulnar arteria continuous circulatory compression using a customized pulse wave sphygmomanometer on the basis of the control group for a duration of 4 hours. Puncture site bleeding, pain, thumb skin temperature, nursing workload, delayed pressure measurement rate, radial artery blood flow velocity, and rates of radial artery occlusion and stenosis were compared between the two groups 24 hours postoperatively. Results There were no significant differences in puncture site bleeding, pain, thumb skin temperature, nursing workload, and delayed pressure measurement rate between the two groups (P>0.05). However, the experimental group presented significantly faster radial artery blood flow velocity 0.5 hours before removing the blood pressure monitor, as well as lower rates of radial artery occlusion and stenosis 24 hours postoperatively compared to the control group (P < 0.05). Conclusion The use of a blood pressure monitor for selective continuous circulatory compression for a duration of 4 hours can enhance radial artery blood supply and reduce rates of postoperative radial artery occlusion and stenosis in elderly coronary intervention patients under the premise of not increasing the risk of bleeding, pain, nurse workload and delayed pressure extraction.

Esophageal squamous cell carcinoma (ESCC) is one of the leading causes of cancer death worldwide. It is urgent to develop new drugs to improve the prognosis of ESCC patients. Here, we found benzydamine, a locally acting non-steroidal anti-inflammatory drug, had potent cytotoxic effect on ESCC cells. Benzydamine could suppress ESCC proliferation in vivo and in vitro. In terms of mechanism, CDK2 was identified as a target of benzydamine by molecular docking, pull-down assay and in vitro kinase assay. Specifically, benzydamine inhibited the growth of ESCC cells by inhibiting CDK2 activity and affecting downstream phosphorylation of MCM2, c-Myc and Rb, resulting in cell cycle arrest. Our study illustrates that benzydamine inhibits the growth of ESCC cells by downregulating the CDK2 pathway.
Humans ; Benzydamine ; Esophageal Neoplasms/drug therapy* ; Esophageal Squamous Cell Carcinoma/drug therapy* ; Molecular Docking Simulation ; Phosphorylation ; Cell Proliferation ; Cell Line, Tumor ; Apoptosis ; Cyclin-Dependent Kinase 2

Gene therapy is a rapidly developing field. The most widely used technique for foreign gene transfer is lentiviral-mediated gene therapy. Lentiviral vector has been developed from the first generation to the third generation in terms of safety. The preparation of lentiviruses with high titer remains difficult. In this study, a Fibra-Cel sheet carrier was used as an HEK293T cell carrier matrix, and several sterile cell culture spinners were combined and cultured on a roller bottle machine to scale up the adherent cells. The virus titer was maximized by screening the factors to optimize the lentivirus titer in the third-generation lentivirus packaging process one by one. Fibra-Cel sheet vector was successfully used as the matrix of HEK293T cell adhesion to culture adherent cells at large scale. The optimal conditions for large-scale preparation of the third-generation lentivirus by bottle roller were screened and three batches of lentiviruses were produced on pilot scale. The production time of lentivirus was shortened from 120 hours to 54 hours from plasmid transfection to virus collection; in terms of cost, a rolling bottle machine was used instead of a bioreactor, leading to lower cost and no need for repeated sterilization during the whole process. The safe, effective and low-cost operation of successful production will provide a technical base for the large-scale preparation of lentivirus and thus lay a firm foundation for its clinical application.
Genetic Vectors ; HEK293 Cells ; Humans ; Lentivirus ; Transduction, Genetic ; Transfection

Cancer is a common cause of death worldwide. Despite significant advances in cancer treatments, the morbidity and mortality are still enormous. Tumor heterogeneity, especially intratumoral heterogeneity, is a significant reason under-lying difculties in tumor treatment and failure of a number of current therapeutic modalities, even of molecularly targeted therapies. The development of a virtually noninvasive“liquid biopsy”from the blood has been attempted to characterize tumor heterogeneity. This review focuses on cell-free circulating tumor DNA (ctDNA) in the bloodstream as a versatile biomarker. ctDNA analysis is an evolving field with many new methods being developed and optimized to be able to successfully extract and analyze ctDNA, which has vast clinical applications. ctDNA has the potential to accurately genotype the tumor and identify personalized genetic and epigenetic alterations of the entire tumor. In addition, ctDNA has the potential to accurately monitor tumor burden and treatment response, while also being able to monitor minimal residual disease, reducing the need for harmful adjuvant chemotherapy and allowing more rapid detection of relapse. There are still many challenges that need to be overcome prior to this biomarker getting wide adoption in the clinical world, including optimization, standardization, and large multicenter trials.

OBJECTIVETo explore the effects of acrylonitrile on T lymphocyte subsets, expression of toll-like receptor 4 and related cytokines in rats.

METHODSSixty-four Sprague-Dawley rats were randomly divided into 4 female groups and 4 male groups, and there were 8 rats in each group. Rats in each group were respectively given a single dose of 0, 5, 10 and 20 mg/kg acrylonitrile by gavage, once a day, 5 days a week, for 13 weeks. Blood and spleen T lymphocyte subsets was detected by flow cytometry, the mRNA expression of TLR4, IL-1β and TNF-α was analyzed by real-time quantitative PCR, the protein expression of TLR4 was evaluated by Western blot.

RESULTSCompared with control group, the percentages of blood CD3, CD4 T cells in 20 mg/kg female group and CD4/CD8 ratio in 5, 10 and 20 mg/kg female groups was significantly decreased, CD8 T cells in 20 mg/kg group was significantly increased (P < 0.05 or P < 0.01), blood CD3 T cells in 5 mg/kg male group, CD4 T cells and CD4/CD8 ratio in 20 mg/kg male groups were lower than that of control group, CD8 T cells in 20 mg/kg make group was significantly in oreased (P < 0.05 or P < 0.01). Spleen CD4, CD8 T lymphocyte percentages and CD4/CD8 ratio in 20 mg/kg female group decreased significantly, CD8 T cells in 20 mg/kg male group was significantly increased (P < 0.05 or P < 0.01), spleen CD3, CD4, CD8 T cells in 20 mg/kg male group and CD4/CD8 ratio in 10, 20 mg/kg male groups was also significantly decreased, CD3 T cells in 20 mg/kg and CD8 T cells in 10, 20 mg/kg male groups were significantly increased (P < 0.05 or P < 0.01) (TLR4 mRNA was lower expressed in 5, 10 and 20 mg/kg male groups and 10 mg/kg female group (P < 0.05 or P < 0.01), and TLR4 protein in 5 mg/kg female group and 20 mg/kg male group was significantly lower than control group (P < 0.05). The expression level of IL-1β mRNA was significantly decreased in 5, 10 and 20 mg/kg female group and 5, 10 mg/kg male group (P < 0.05 or P < 0.01), TNF-α mRNA was lower expressed in 10, 20 mg/kg female groups and 5, 10 mg/kg male groups (P < 0.01).

CONCLUSIONAcrylonitrile may lead to the changes of CD3, CD4, CD8 T lymphocyte percentages and CD4/CD8 ratio in rat blood and spleen, and also significantly effected the expression level of TLR4 mRNA and protein together with the secretion of IL-1β, TNF-α. This may cause effects on the cellular immune function.

Acrylonitrile ; toxicity ; Animals ; Female ; Interleukin-1beta ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; T-Lymphocyte Subsets ; drug effects ; immunology ; Toll-Like Receptor 4 ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

Objective To investigate the safety and efficacy of intravenous thrombolytic therapy with recombinant tissue plasminogen activator(rtPA)in patients with isolated penetrating artery territory infarct (IPAI).Methods Data of retrospectively collected clinical,laboratory,and radiological from 75 consecutive patients with acute ischemic stroke treated with intravenous rtPA therapy from June 2009 to April 2011.Etiological classification was carried out according to the Chinese Ischemic Stroke Classification of Subgroups(CISS).The rates of hemorrhagic transformation(HT)and clinical outcomes of patients were compared between IPAI group and non-IPAI group.Results All 75 patients with mean age of 67.4years and 25(33.3％)fenale,were treated with intravenous rtPA.Before treatment,their average score of the National Institutes of Health Stroke Scale(NIHSS)was 12.3 ± 6.4,and mean length of time from onset to treatment was 239.6 ±97.5 minutes.After thrombolytic therapy,the radiological HT was found in 24 patients(32％).Symptomatic intracraneal hemorrhage(ICH)occurred in 4 patients(5.3％).Of 22 (29.3％)patients with IPAI,only one experienced HT.Logistic regression analysis suggested that IPAI wasan individualized predictor used alone for determining the low risk of HT.In the patients with IPAI,82％ of them had an individual clinical outcome(mRS ＜ 2)one month after onset,and the neurological outcomes were better in patients with IPAI than those in patients with non-IPAI(P ＜ 0.01).Conclusions The risk of hemorrhagic complication was low and the clinical outcome was good in patients with isolated penetrating artery territory infarct after intravenous thrombolytic therapy with rtPA.Imaging diagnosis of IPAI might facilitate the treatment with rtPA in this cohort of patients.