Objective To establish U251 cells with inhibited expression of interferon-γ inducible protein 30 (IFI30), and to investigate the effect of IFI30 on cell biological function as well as its underlying mechanism. Methods Three knockdown sequences which target IFI30 were designed online and 3 small interfering RNAs (siRNA) were synthesized. After transfection, the inhibition efficiency was detected by real-time quantitative PCR. The siRNA sequence with the highest inhibition efficiency was selected to create short hairpin RNA (shRNA) plasmids. The recombinant plasmids and packaging plasmids were co-transfected into HEK293T cells to prepare lentivirus. The glioma U251 cells were transfected with lentivirus, and the positive cells were screened by puromycin. CCK-8 assay, 5-ethyl-2'-deoxyuridine (EdU) and colony formation assays were used to analyze cell proliferation; the flow cytometry was used to analyze cell cycle and apoptosis; the Transwell^TM assay was used to detect cell invasion; the wound-healing assay was employed to detect cell migration, and western blot analysis to detect the protein expresison of cyclin D1, B-cell lymphoma factor 2 (Bcl2), epithelial cadherin (E-cadherin), neural cadherin (N-cadherin), signal transducer and activator of transcription 1 (STAT1). Results The sequence which effectively target IFI30 was screened and U251 cell line capable of inhibiting the IFI30 expression was successfully established. When IFI30 expression was knocked down, the proliferation of U251 cells was inhibited, along with increased ratio of cells in the phase G0/G1, the decreased phase S, the increased rate of cell apoptosis. The cell invasion and migration capabilities was also reduced. The decreased expression of cyclin D1, Bcl2 and N-cadherin were observed in U251 cells, and the expression of E-cadherin and the phosphorylation of STAT1 were found increased. Conclusion Knockdown of IFI30 inhibits the proliferation, invasion and migration of human glioma cell U251 and promotes its apoptosis by activating STAT1.
Humans ; Cyclin D1/genetics* ; HEK293 Cells ; Interferon-gamma ; RNA, Small Interfering ; Apoptosis/genetics* ; Cadherins ; Cell Proliferation/genetics* ; Glioma/genetics* ; Proto-Oncogene Proteins c-bcl-2 ; Oxidoreductases Acting on Sulfur Group Donors ; STAT1 Transcription Factor/genetics*

Objective To establish a novel real-time quality control method for rapidly identifying the random error of sodium ion con-centration in serum using an artificial intelligence voting algorithm,and evaluate the relevant effectiveness of the model established on this basis.Methods A total of 144 754 test results of serum sodium ion rom the inpatients measured by Beckman AU5400 biochemis-try analyzer from January to May 2021 were obtained retrospectively from laboratory information system of the Department of Clinical La-boratory,Nanjing Drum Tower Hospital,and all the data were used as unbiased data for the current study.The random errors were arti-ficially introduced to generate the corresponding biased data set.Subsequently,the voting algorithm-based internal quality control model(ViQC)was established using the principles of the voting algorithm.The ViQC model and five classical PBRTQC(patient-based real-time quality control)algorithms were performed direct to each biased data.The analytical performance of the ViQC model was evaluated by using classification model criteria.The trimmed average number of patient samples until error detection(tANPed)was used to com-pare the clinical detection efficacy of the ViQC model with those of the five classical algorithms,and the error detection curves were plotted.Results Compare with all the classical algorithms,the ViQC model showed a false positive rate below 0.002 and achieved ac-curacy above 0.951 in detecting all the deviations.When the error factors were 1.5,2.5,and 3.0,the false positive rate of the ViQC model was zero.When the error factor was 2.5,its accuracy reached 0.979.Compared to the five classical PBRTQC algorithms,the ViQC model reduced the overall average tANPed by up to 34％and showed higher sensitivity for error detection.In addition,the ViQC model demonstrated the area under the ROC curve was as high as 0.989 at TEa on the test set,but the value of tANPed wasonly five.Conclusion We successfully established a real-time quality control model for the data of patients based on artificial intelligence algo-rithms,and its efficacy of clinical detection was superior to the traditional PBRTQC algorithms.

Objective To investigate the influence of oral midazolam on postoperative behavioral changes(NPOBCs)in preschool children undergoing laparoscopic inguinal hernia sac ligation.Methods A total of 197 preschool children,118 males and 79 females,aged 1-6 years,BMI 14.0-24.5 kg/m2,ASA physical status Ⅰ or Ⅱ,who undergoing laparoscopic inguinal hernia sac ligation from July 2022 to A-pril 2023 were selected.The children were divided into two groups by random number table method:the oral midazolam group(group M,n=100)and the oral saline group(group C,n=97).In the observation room,group M received an oral dose of 0.5 mg/kg midazolam while group C received an oral dose of 5％sa-line 0.25 ml/kg before being accompanied by a clown doctor to the operating room after 30 minutes.Modi-fied Yale preoperative anxiety scale(m-YPAS)scores before entering the observation room and before ente-ring the operating room,as well as induction compliance checklist(ICC)scores before anesthesia induction were recorded.Postoperatively,FLACC and PAED scores were recorded at immediately,6,and 24 hours after surgery along with occurrences of postoperative nausea and vomiting and hypoxemia.Family members of the children were followed up at 1 day,7,14,and 30 days after discharge using PHBQ to record NPOBCs occurrence in children.Results Compared with group C,m-YPAS score,ICC score before anesthesia in-duction,PAED score immediately after surgery,and the incidence of NPOBCs 1 day,7,14,and 30 days after surgery were significantly reduced in group M(P＜0.05).There were no significant differences in YPAS scores before entering the observation room,6 and 24 hours PAED scores,incidence of postoperative nausea and vomiting,hypoxemia between the two groups.Conclusion Preoperative oral administration of midazolam to children undergoing laparoscopic inguinal hernia sac ligation can effectively alleviate periopera-tive anxiety and reduce the incidence of NPOBCs.It does not increase the risk of delayed awakening or post-operative adverse reactions in children.

This research aimed to explore strategies to enhance the influence of major themed publicity in hospital party building within several prominent tertiary public hospitals in China,including one in Beijing,one in Zhejiang and four in Guang-dong.An analysis of themed publicity initiatives over the past five years,particularly during pivotal party events and key thematic activities,was undertaken.This assessment included both offline events and the online media coverage through various integrated media platforms.The study aimed to offer development suggestions for hospitals to amplify the influence of their major themed publicity strategies for Party building.Currently,hospitals are placing increased emphasis on the strategic planning of major themed publicity for Party building.Innovation in content and promotional tactics,along with strengthened integration with main-stream media,is essential.Effective dissemination that elicits empathy and presents high-quality publicity materials is crucial for achieving optimal dissemination outcomes and further elevating the hospital's Party building influence.

Objective:To observe the effects of sacubitril/valsartan (LCZ696) on viral replication and cardiomyocyte apoptosis in mice with coxsackievirus B3 (CVB3)-induced viral myocarditis (VMC) and to analyze the underlying mechanisms.Methods:Forty BALB/c mice were randomly divided into four groups with 10 in each group: Sham, Sham+ LCZ696, VMC, and VMC+ LCZ696 groups. VMC model was established by intraperitoneal injection of 0.1 ml of CVB3 with a concentration of 10 6 TCID 50/ml into BALB/c mice, while the sham intervention was an equal volume of saline. The day of virus injection was defined as day 0. LCZ696 was administered by gavage at a dose of 60 mg/kg every day for seven consecutive days starting from day 1. Mouse survival rates were calculated. Echocardiography was used to evaluate the cardiac function of mice. The level of creatine kinase-MB (CK-MB) was detected by ELISA. Western blot was used to detect the levels of inflammatory cytokines (IL-6, TNF-α), apoptosis-related proteins (caspase-3, cleaved-caspase-3, Bax, Bcl-2), CVB3 surface protein (VP-1) and p-AKT/AKT in the hearts of mice. CVB3 mRNA in mouse hearts was measured by PCR. Inflammatory cell infiltration and cell apoptosis in mouse hearts were observed by HE staining and TUNEL staining, respectively. Results:Compared with the Sham group, the mice in the VMC group had a decreased survival rate and impaired cardiac function ( P<0.05). The levels of CK-MB, IL-6, TNF-α, cleaved-caspase-3/caspase-3, Bax/Bcl-2, VP-1, and CVB3 mRNA in the hearts of VMC mice increased significantly ( P<0.05), accompanied by increased expression of AKT, decreased phosphorylation of AKT ( P<0.05) and increased cell apoptosis. LCZ696 reversed the above changes. It could increase the survival rate, improve the cardiac function ( P<0.05), decrease cardiac inflammation, cell apoptosis and viral replication ( P<0.05), and increase the phosphorylation of AKT ( P<0.05). LCZ696 had no significant effects on the survival rate, cardiac function, myocardial injury, cardiac inflammation, cell apoptosis, viral replication or the expression of PI3K/AKT signaling pathway-related proteins in normal mice. Conclusions:LCZ696 could significantly inhibit cardiomyocyte apoptosis and reduce CVB3 replication in the hearts of VMC mice by regulating the PI3K/AKT pathway, thereby improving mouse cardiac function and survival rate.

OBJECTIVE To identify the chemical components of Changtong oral liquid （CTOL），and to provide reference for the basic research and secondary development of its pharmacological substances. METHODS UHPLC-Orbitrap HRMS technique was adopted. CTOL sample was separated on a Hypersil Gold column with mobile phase consisted of 0.1% formic acid （containing 5 mmol/L ammonium formate）-acetonitrile （gradient elution）. The eluent was detected in positive and negative ion modes using an electrospray ionization source. The data was processed by Xcalibur 4.3 and Compound Discoverer 3.3 software. The primary and secondary mass spectra data of each compound were collected. The unknown compounds were identified according to the mass spectrometry library of the instrument and the network databases mzCloud，mzVault，etc. Through matching with the pharmacology database and analysis platform of the traditional Chinese medicine system，the chemical components could be attributed to the traditional Chinese medicine. RESULTS Fifty-three chemical components were identified and analyzed from CTOL，such as 24 flavonoids，8 quinones，5 phenylpropanoids，4 sugars and glycosides，5 organic acids，3 amino acids，1 alkaloid，1 phenolic and 2 other compounds. Among them，12 components were derived from Salvia miltiorrhiza，9 from Citrus aurantium，7 from Rheum palmatum，4 from Angelica sinensis，1 from Magnolia officinalis，16 from Glycyrrhiza uralensis，and 4 from many kinds of medicinal materials. CONCLUSIONS CTOL mainly contains flavonoids，quinones and phenylpropanoid compounds，and its chemical components mainly come from G. uralensis，S. miltiorrhiza and C. aurantium.

OBJECTIVE: To investigate the growth-inhibitory and pro-apoptotic effects of piroctone olamine (PO) on glioma cells and explore the underlying mechanism. METHODS: Human glioma cell lines U251 and U373 were treated with PO and the changes in cell proliferation were detected using CCK-8 assay and EdU assay. Clone formation assay and flow cytometry were used to examine the changes in clone formation ability and apoptosis of the treated cells. Mitochondrial membrane potential of the cells and morphological changes of the mitochondria were detected using JC-1 staining and a fluorescence probe, respectively. The expressions of mitochondrial fission protein DRP1 and the fusion protein OPA1 were determined with Western blotting. Transcriptome sequencing and differential gene enrichment analysis was performed, and the expression levels of PI3K, AKT and p-AKT in the treated cells were verified using Western blotting. RESULTS: CCK-8 assay showed that PO significantly inhibited the proliferation of U251 and U373 cells in a time- and dose-dependent manner (P < 0.001). EdU test showed that the proliferative activity of PO-treated cells was significantly decreased, and the number of cell colonies also decreased significantly (P < 0.01). PO treatment significantly increased apoptotic rates (P < 0.01), decreased mitochondrial membrane potential and caused obvious changes in mitochondrial morphology of the cells. Pathway enrichment analysis showed that the down-regulated genes were significantly enriched in the PI3K/AKT pathway, which was verified by Western blotting showing significantly down-regulated expression levels of PI3K, AKT and p-AKT in PO-treated cells (P < 0.05). CONCLUSION PO interferes with mitochondrial fusion and fission function through the PI3K/AKT pathway, thereby inhibiting the proliferation and increasing apoptosis of glioma cells.
Humans ; Glioma ; Mitochondrial Dynamics ; Phosphatidylinositol 3-Kinases ; Proto-Oncogene Proteins c-akt

OBJECTIVE: To investigate the effect of lactic acid-induced upregulation of PLEKHA4 expression on biological behaviors of glioma cells and the possible molecular mechanism. METHODS: GEO database and GEPIA2 website were used to analyze the relationship between PLEKHA4 expression level and the pathological grade of glioma. A specific PLEKHA4 siRNA was transfected in glioma U251 and T98G cells, and the changes in cell proliferation ability were assessed by real-time cell analysis technology and Edu experiment. The colony-forming ability of the cells was evaluated using plate cloning assay, and cell cycle changes and cell apoptosis were analyzed with flow cytometry. The mRNA expression of PLEKHA4 was detected by PCR in glioma samples and controls and in glioma cells treated with lactic acid and glucose. Xenograft mice in vivo was used to detect tumor formation in nude mice; Western blotting was used to detect the expressions of cyclinD1, CDK2, Bcl2, β-catenin and phosphorylation of the key proteins in the MAPK signaling pathway. RESULTS: The results of GEO database and online website analysis showed that PLEKHA4 was highly expressed in glioma tissues and was associated with poor prognosis; PLEKHA4 knockdown obviously inhibited the proliferation and attenuated the clone-forming ability of the glioma cells (P < 0.05). Flow cytometry showed that PLEKHA4 knockdown caused cell cycle arrest in G1 phase and promoted apoptosis of the cells (P < 0.01). PLEKHA4 gene mRNA expression was increased in glioma samples and glioma cells after lactate and glucose treatment (P < 0.01). PLEKHA4 knockdown, tumor formation ability of nude mice decreased; PLEKHA4 knockdown obviously lowered the expression of cyclinD1, CDK2, Bcl2 and other functional proteins, inhibited the phosphorylation of ERK and p38 and reduced the expression of β-catenin protein (P < 0.01). CONCLUSION PLEKHA4 knockdown inhibited the proliferation of glioma cells and promoted apoptosis by inhibiting the activation of the MAPK signaling pathway and expression of β-catenin. Lactic acid produced by glycolysis upregulates the expression of PLEKHA4 in glioma cells.
Humans ; Animals ; Mice ; Up-Regulation ; beta Catenin/metabolism* ; Mice, Nude ; Brain Neoplasms/pathology* ; Lactic Acid ; Cell Line, Tumor ; Glioma/pathology* ; Cell Proliferation ; Apoptosis ; Proto-Oncogene Proteins c-bcl-2/metabolism* ; RNA, Messenger/genetics* ; Gene Expression Regulation, Neoplastic

Objective:To analyze the difference between Z score and previous criteria in the diagnosis characteristics of coronary artery aneurysm (CAA) in Kawasaki disease, and to investigate the clinical distribution of Kawasaki disease CAA in the Z score group.Methods:This study retrospectively analyzed the clinical and echocardiographic data of 2 419 children with Kawasaki disease in Shenzhen Children′s Hospital from January 2009 to December 2019. The traditional criteria and Z score criteria were used to diagnose CAA, and the differences of diagnostic efficiency between the 2 diagnostic methods were analyzed. The clinical distribution characteristics of CAA in children with Kawasaki disease were analyzed by grouping their sex, clinical classification (complete Kawasaki disease, incomplete Kawasaki disease) the sensitivity to intravenous immunoglobulin (IVIG) (IVIG-sensitive Kawasaki disease,IVIG-unresponsive Kawasaki disease). And the course of the disease (≤6 weeks, >6-8 weeks, >8 weeks to 6 months) etc. The χ2 test or Kruskal-Wallis test was used for comparison between the groups, and the Kappa test was used for consistency evaluation.Results:Among the 2 419 children with Kawasaki disease, 1 558 were males and 861 were females. The age of onset was 1.8 (1.0, 3.2) years. The rate of CAA by Z score criteria was higher than that by traditional method (21.9% (529/2 419) vs. 13.9% (336/2 419), χ 2=1 074.94, P<0.001). Compared to the traditional method, the Z score criteria found higher rate of CAA in male patients, patients with incomplete Kawasaki disease, and IVIG-unresponsive patients (25.2% (392/1 558) vs. 16.0% (249/1 558), (32.7% (166/507) vs. 19.5% (99/507), 30.5% (95/312) vs. 24.0% (75/312), χ 2=694.05, 216.19, 184.37, all P<0.001). The Z score criteria was consistent with the traditional method in diagnosing CAA (κ=0.642, P<0.001). Moreover, in the Z score criteria, the rate of CAA in males (25.2%, 392/1 558) was higher than that in females (15.9%, 137/861), higher in incomplete Kawasaki cases (32.7%, 166/507) than that in complete Kawasaki case (19.0%, 363/1 912), and higher in IVIG-unresponsive cases (30.4%, 95/312) than that in IVIG-sensitive cases (20.6%, 434/2 107), with statistically significant differences (χ 2=27.76, 44.38, 15.43, all P<0.001). Coronary Z score of course ≤ 6 weeks was greater than that of course between>6-8 weeks and >8 weeks to 6 months (1.3 (0.7, 2.3) vs. 0.7 (0.3, 1.4), 0.7 (0.3, 1.3), Z=20.65, 13.70, both P<0.001). Conclusions:The rate of CAA in Kawasaki disease by Z score criteria is higher than that by traditional method. In the Z score group, most CAA occur within 6 weeks of the course of the disease, and the rate of CAA in male, incomplete Kawasaki disease, and IVIG-unresponsive is higher.

Objective:At present, there are several modes of patent transformation of hospital service invention in our country. However, most of them are one-to-one style which is time-consuming and difficult to achieve industrialization in a short time. This study focuses on whether it is appropriate for public hospitals to use the mode of general license (i.e. one patent licensed to several enterprises concurrently or successively) to turn the patents of invention into productivity more efficiently.Methods:Combined with the period of COVID-19 pandemic, Zhongshan Hospital Fudan University successfully applied the mode of general license to achieve the patent transfer experience, and discussed the important value of the mode of general license for translational medicine.Results:The latest achievements of medical research are turned into clinical medical technologies and products quickly and effectively, proposing an appropriate scheme for the rapid transformation of patents, which balanced the interest of different stakeholders to gain the win-win situation.Conclusions:The innovative general licensing model can enable the patents transformation quickly and efficiently as long as it′s used properly.