1. A new strategy for evaluating antitumor activity in vitro with time-dimensional characteristics of RTCA technology
Fang-Tong LIU ; Shu-Yan XING ; Jia YANG ; Guo-Ying ZHANG ; Rong RONG ; Xiao-Yun LIU ; Dong-Xue YE ; Yong YANG ; Xiao-Yun LIU ; Dong-Xue YE ; Rong RONG ; Yong YANG ; Xiao-Yun LIU ; Dong-Xue YE ; Yong YANG ; Xiao-Yun LIU ; Dong-Xue YE ; Yong YANG
Chinese Pharmacological Bulletin 2024;40(3):592-598
Aim To analyze the anti-A549 and HI299 lung ade-nocarcinoma activities via using examples of baicalin, astragalo-side, hesperidin and cisplatin based on real time cellular analysis (RTCA) technology, and to build a new strategy for EC50 e-valuation reflecting the time-dimensional characteristic. Methods Using RTCA Software Pro for data analysis and GraphPad Prism and Origin Pro plotting, the in vitro anti-A549 and H1299 lung adenocarcinoma activities of baicalin, astragaloside, hesperidin, and cisplatin were characterized using the endpoint method and time dimension, respectively. Results (X) There were significant differences in EC50 values of A549 and H1299 cells at 24 h and 48 h endpoint methods. (2) The correlation coefficient of the curve fitted with the four-parameter equation was > 0. 9, and the dynamic change of EC50 remained relatively stable (the linear fitting of EC50 at adjacent 4 points I slope 1^1) used to calculate the EC50 value within this time dimension. The EC50 of baicalin, astragaloside, hesperidin and cisplatin on A549 cells was 52. 97 ±1.75 плпо! • L~1(16~48 h) , 62.88 ± 2.91 ijunol • L"1 (32.25 -48 h) , 78.84 ±0.33 плпо1 • L"1 (21.5 -29.75 h), 13.57 ±1.54 плпо1 • L_1(27.5 -48 h), respectively; the EC50 of baicalin, astragaloside, hesperidin and cisplatin on H1299 cells was 43. 71 ± 1. 26 |лто1 • L_1 ( 19. 5 -48 h), 47.23 ±1. 19 |лто1 • L_1(14 -48 h) , 39.45 ±0.24 плпо1 • L"1 (12.75 -46.25 h), 25.97 ±4.76 плпо1 • L"1 (10. 25 -48 h) , respectively. The results showed that the time window for the anti-tumor effect of the test solution/drug was different. Conclusions Based on RTCA technology, it is more accurate and reasonable to select EC50 data that exhibit better fitting, stable changes, and time-dimensional characteristics for the evaluation of anti-tumor activity. In addition, this method of distinguishing different effective time of antitumor drugs can provide a reference for the timing of clinical combination drugs, and this approach will also provide a reference for further related studies.
2.Bibliometric and visual analysis of Chinese scarlet fever literature
Chunyu ZHAO ; Liu LONG ; Xinjing JIA ; Chunyuan DUAN ; Lisha LIU ; Xiushan ZHANG ; Jinpeng GUO ; Ruizhong JIA ; Wenyi ZHANG ; Yong WANG
Journal of Public Health and Preventive Medicine 2024;35(2):1-5
Objective To analyze the research status and trend of scarlet fever literature in China, and to provide reference for subsequent research. Methods Three major Chinese databases, CNKI, Wanfang, and VIP, as well as Web of Science English database, were used to search for literature related to scarlet fever from 2000 to 2023. Citespace6.2.R2 software was used to statistically analyze the number of publications, authors, institutions and journals, co-cited literature, keyword clustering, and other literature characteristics of the literature. Results From 2000 to 2023, a total of 1 011 Chinese literature were included in the three major Chinese databases. Since 2011, the number of publications had gradually increased, but in recent years, the number of publications had decreased. The organization with the most publications was the Shenyang Center for Disease Control and Prevention. The cluster analysis of key words mainly formed 9 cluster tags, and the high-frequency keywords mainly included epidemic characteristics, epidemiology, incidence rate, etc. A total of 84 English literature were included in the WOS database, with an overall upward trend in publication volume. The institution with the most publications was the China Center for Disease Control and Prevention, and the most frequently cited journal was “LANCET INFECT DIS”.《Resurgence of scarlet fever in China: a 13-year population-based surveillance study》 was the most cited journal. After keyword cluster analysis, 9 cluster labels were mainly formed, and the keywords were mainly outbreak,Hong Kong, and Group A streptococcus. Conclusion Compared with the English literature, which mainly focuses on spatiotemporal aggregation, etiology and strain resistance, Chinese literature focuses more on epidemic surveillance, clinical features and quality nursing.
3.Analysis of phenotype formation mechanism of a new variety of Lonicera japonica Flos "Huajin 6" at long bud stage
Run-zhu LI ; Cong-lian LIANG ; Zhen-hua LIU ; Jia LI ; Yong-qing ZHANG ; Hai-yan LIU ; Gao-bin PU
Acta Pharmaceutica Sinica 2024;59(2):476-481
Based on the long bud stage phenotype of a new
4.Genomic characteristics analysis of a colistin and tigecycline-resistant Klebsiella pneumoniae
Xinjing JIA ; Xinran GONG ; Peng LI ; Chuanyuan DUAN ; Lisha LIU ; Dayang ZOU ; Yong WANG
Journal of Public Health and Preventive Medicine 2024;35(3):37-41
Objective In this study, a strain of colistin and tigecycline-resistant bacteria isolated in 2009 was analyzed, and the structure of drug-resistant plasmid and genetic environment were discussed, so as to provide basis for the prevention and control of multidrug-resistant bacteria. Methods A strain (GZ12244) with positive mcr and tet(M) was obtained by screening colistin and tigecycline resistance genes. Vitek-2 was used for strain identification, and the drug sensitivity test was carried out by broth dilution method. The molecular typing, drug resistance genes, insertion sequences, plasmid structure and genetic background were analyzed by genome-wide sequencing and bioinformatics. Results Strain GZ12244 is Klebsiella pneumoniae, which is resistant to colistin B, tigecycline, cefuroxime and tetracycline, and carries a variety of drug-resistant related genes such as mcr-1 and tet(M), and some of the drug-resistant genes with antibiotic efflux and antibiotic target change have amino acid substitution mutations. Mcr-1 and tet(M) coexist in a plasmid, and mcr-1 flanked by two insertion sequences ISApl1. There are insertion sequences such as IS15, IS1D and ISEc63 in the upstream and downstream of tet(M) gene. Conclusion Klebsiella pneumoniae GZ12244 is a multidrug-resistant strain. The drug-resistant gene exists in plasmid, and the mobile elements in upstream and downstream may spread the drug-resistant gene.
5.Forensic Identification and Evaluation of 25 Obstetric Brachial Plexus Palsy Medical Damage Cases
Yong YU ; Ying-Jie WANG ; Yun-Fei JIA ; Bao-Jing HUANG ; Song-Yue HE ; Chuan-Chuan LIU
Journal of Forensic Medicine 2024;40(1):43-49
Objective To analyze the high risk factors of obstetric brachial plexus palsy(OBPP),and to explore how to evaluate the relationship between fault medical behavior and OBPP in the process of medical damage forensic identification.Methods A retrospective analysis was carried out on 25 cases of medical damage liability disputes related to OBPP from 2017 to 2021 in Beijing Fayuan Judicial Science Evidence Appraisal Center.The shortcomings of hospitals in birth weight assessment,delivery mode selection,labor process observation and shoulder dystocia management,and the causal relation-ship between them and the damage consequences of the children were summarized.Results Fault medi-cal behavior was assessed as the primary cause in 2 cases,equal cause in 10 cases,secondary cause in 8 cases,minor cause in 1 case,no causal relationship in 1 case,and unclear causal force in 3 cases.Conclusion In the process of forensic identification of OBPP,whether medical behaviors fulfill diagno-sis and treatment obligations should be objectively analyzed from the aspects of prenatal evaluation,de-livery mode notification,standardized use of oxytocin,standard operation of shoulder dystocia,etc.Meanwhile,it is necessary to fully consider the objective risk of different risk factors and the diffi-culty of injury prevention,and comprehensively evaluate the causal force of fault medical behavior in the damage consequences.
6.Clinical analysis of aortographic manifestations in 30 child patients with Kawasaki disease
Jia FU ; Yong ZHANG ; Jing LIU ; Yong YANG ; Aiguo ZHAI ; Juanjuan DING
Journal of Interventional Radiology 2024;33(2):126-129
Objective To summarize the aortographic features of child patients with Kawasaki disease(KD)complicated by coronary artery lesion(CAL)who were treated at the Wuhan Children's Hospital of China.Methods The clinical data of 30 child patients with early recovery stage KD complicated by CAL,who received aortic angiography examination at Wuhan Children's Hospital of China between June 2016 and June 2023,were retrospectively analyzed.Preoperative echocardiography was adopted to evaluate the severity of CAL,and aortic angiography under laryngeal mask general anesthesia was performed.Results Successful aortic angiography examination was accomplished in all the 30 child patients with KD complicated by CAL.After angiography,the CAL risk classification results showed that grade I was seen in 5,grade Ⅱ in 7,grade Ⅲ in 12,grade Ⅲ in 4,and grade V in 2 child patients.Of all child patients,old coronary thrombosis was found in one(grade Ⅲb),and coronary artery stenosis with collateral circulation formation in one(grade Ⅴ).The CAL was located at the proximal segment(n=15),middle segment(n=3)and distal segment(n=2)of the right coronary artery,at the left main coronary artery(n=12),and at the proximal segment of left coronary artery anterior descending branch(n=9).Electrocardiography showed sinus rhythm(n=17),sinus arrhythmia(n=4),right bundle branch block(n=4),atrial premature beat(n=1),ventricular premature beat(n=2),T-wave change(n=1),and Q-wave change(n=1,coronary lesion grade V,echocardiography suggested left heart enlargement).No cardiac enlargement,cardiac functional insufficiency,or valvular regurgitation signs were observed in the remaining child patients.Conclusion It is clinically safe and feasible to perform aortic angiography examination in child patients with early recovery stage KD complicated by CAL.In this series of child patients with KD,CAL mainly occurs at the proximal segment of coronary artery,especially at the proximal segment of right coronary artery as well as at the left main artery and left anterior descending branch.
7.PRMT7 Regulates Adipogenic Differentiation of hBMSCs by Modulating IGF-1 Signaling
Qian GUO ; Jia QING ; Da-Zhuang LU ; Xu WANG ; Yang LI ; Hui ZHANG ; Ying-Fei ZHANG ; Yun-Song LIU ; Yong-Sheng ZHOU ; Ping ZHANG
Progress in Biochemistry and Biophysics 2024;51(6):1406-1417
ObjectiveProtein arginine methyltransferases (PRMTs) play pivotal roles in numerous cellular biological processes. However, the precise regulatory effects of PRMTs on the fate determination of mesenchymal stromal/stem cells (MSCs) remain elusive. Our previous studies have shed light on the regulatory role and molecular mechanism of PRMT5 in MSC osteogenic differentiation. This study aims to clarify the role and corresponding regulatory mechanism of PRMT7 during the adipogenic differentiation of bone marrow-derived mesenchymal stem cells (BMSCs). Methods(1) Human bone marrow-derived mesenchymal stem cells (hBMSCs) were cultured in a medium that induces adipogenesis. We used qRT-PCR and Western blot to monitor changes in PRMT7 expression during adipogenic differentiation. (2) We created a cell line with PRMT7 knocked down and assessed changes in PRMT7 expression and adipogenic capacity using Oil Red O staining, qRT-PCR and Western blot. (3) We implanted hBMSCs cell lines mixed with a collagen membrane subcutaneously into nude mice and performed Oil Red O staining to observe ectopic lipogenesis in vivo. (4) A cell line overexpressing PRMT7 was generated, and we examined changes in PRMT7 expression using qRT-PCR and Western blot. We also performed Oil Red O staining and quantitative analysis after inducing the cells in lipogenic medium. Additionally, we assessed changes in PPARγ expression. (5) We investigated changes in insulin-like growth factor 1 (IGF-1) expression in both PRMT7 knockdown and overexpressing cell lines using qRT-PCR and Western blot, to understand PRMT7’s regulatory effect on IGF-1 expression. siIGF-1 was transfected into the PRMT7 knockdown cell line to inhibit IGF-1 expression, and knockdown efficiency was confirmed. Then, we induced cells from the control and knockdown groups transfected with siIGF-1 in lipogenic medium and performed Oil Red O staining and quantitative analysis. Finally, we assessed PPARγ expression to explore IGF-1’s involvement in PRMT7’s regulation of adipogenic differentiation in hBMSCs. Results(1) During the adipogenesis process of hBMSCs, the expression level of PRMT7 was significantly reduced (P<0.01). (2) The adipogenic differentiation ability of PRMT7 knockdown group was significantly stronger than that of control group (P<0.001). (3) The ectopic adipogenic differentiation ability of PRMT7 knockdown group was significantly stronger than that of control group. (4) The adipogenic differentiation ability of the PRMT7 overexpression group was significantly weaker than that of the control group (P<0.01). (5) The expression level of IGF-1 increased after PRMT7 knockdown (P<0.000 1). The expression level of IGF-1 decreased after PRMT7 overexpression (P<0.000 1), indicating that PRMT7 regulates the expression of IGF-1. After siIGF-1 transfection, the expression level of IGF-1 in all cell lines decreased significantly (P<0.001). The ability of adipogenic differentiation of knockdown group transfected with siIGF-1 was significantly reduced (P<0.01), indicating that IGF-1 affects the regulation of PRMT7 on adipogenic differentiation of hBMSCs. ConclusionIn this investigation, our findings elucidate the inhibitory role of PRMT7 in the adipogenic differentiation of hBMSCs, as demonstrated through both in vitro cell-level experiments and in vivo subcutaneous transplantation experiments conducted in nude mice. Mechanistic exploration revealed that PRMT7’s regulatory effect on the adipogenic differentiation of hBMSCs operates via modulation of IGF-1 signaling pathway. These collective findings underscore PRMT7 as a potential therapeutic target for fatty metabolic disorders, thereby offering a novel avenue for leveraging PRMT7 and hBMSCs in the therapeutic landscape of relevant diseases.
8.Study on The Mechanism of Sinomenine Hydrochloride Induced Fibroblast Apoptosis in Rabbits with Adhesive Knee Ankylosis
Xin-Ju HOU ; Hong-Feng LEI ; Yong CHEN ; Fu-Xi LI ; Jing-Ning SUN ; Jia-Ming LIU ; Hong-Mei MA
Progress in Biochemistry and Biophysics 2024;51(4):959-968
ObjectiveThis study aimed to observe the impact of sinomenine hydrochloride on the proliferation of fibroblasts and the mRNA expression of related genes in knee joint adhesion and contracture in rabbits. Additionally, we sought to explore its potential mechanisms in combating knee joint adhesion and contracture. MethodsFibroblasts were cultured in vitro, and experimental groups with varying concentrations of sinomenine hydrochloride were established alongside a control group. Cell proliferation was assessed using the CCK-8 assay. Changes in the mRNA expression of fibroblast-related genes following sinomenine hydrochloride treatment were evaluated using RT-qPCR. The impact of the drug on serum levels of inflammatory cytokines was determined using the ELISA method, and the expression of related proteins was assessed using Western blot. ResultsSinomenine hydrochloride was found to inhibit fibroblast viability, with viability decreasing as the concentration of sinomenine hydrochloride increased. The effects of sinomenine hydrochloride in all experimental groups were highly significant (P<0.05). At the mRNA expression level, compared to the control group, sinomenine hydrochloride led to a significant downregulation of inflammatory cytokines in all groups (P<0.05). Additionally, the expression levels of apoptosis-related proteins significantly increased, while Bcl-2 mRNA expression decreased (P<0.05). The mRNA expression levels of the PI3K/mTOR/AKT3 signaling pathway also decreased (P<0.05). At the protein expression level, in comparison to the control group, the levels of inflammatory cytokines IL-6, IL-8, IL-1β, and TGF-β were significantly downregulated in the middle and high-dose sinomenine hydrochloride groups (P<0.05). The expression levels of cleaved-PARP, cleaved caspase-3/7, and Bax increased and were positively correlated with the dose, while the expression levels of the anti-apoptotic protein Bcl-2 and the PI3K/AKT3/mTOR signaling pathway were negatively correlated with the dose. Sinomenine hydrochloride exhibited a significant inhibitory effect on the viability of rabbit knee joint fibroblasts, which may be associated with the downregulation of inflammatory cytokines IL-6, IL-8, and IL-1β, promotion of apoptosis-related proteins cleaved-PARP, cleaved caspase-3/7, and Bax, suppression of Bcl-2 expression, and inhibition of gene expression in the downstream PI3K/AKT3/mTOR signaling pathway. ConclusionSinomenine hydrochloride can inhibit the inflammatory response of fibroblasts in adhesive knee joints and accelerate fibroblast apoptosis. This mechanism may offer a novel approach to improving and treating knee joint adhesion.
9.The effect of ultrasound-guided myofascial tube combined with knee joint posterior capsule space block on postoperative delirium in elderly patients undergoing total knee arthroplasty
Jia LIU ; Xiaoliang WANG ; Wenwen ZHANG ; Yajie XU ; Yong ZHANG ; Zhaojing FANG ; Lili ZHU
Journal of Chinese Physician 2024;26(3):354-359
Objective:To evaluate the effect of ultrasound-guided adductor canal combined with knee joint posterior capsule space block on the recovery period delirium in elderly patients undergoing total knee arthroplasty (TKA).Methods:A total of 68 elderly patients who underwent unilateral TKA at the Nanjing First Hospital from December 2021 to December 2022 were selected. They were randomly divided into a control group ( n=34) and an observation group ( n=34) using a random number table method. Both groups received general anesthesia, and the observation group received ultrasound-guided myofascial tube combined with knee joint posterior capsule space block before the surgery began. The incidence of delirium within 24 hours after surgery was mainly observed in two groups of patients. The surgical time, recovery time, intraoperative blood loss, infusion volume, total use of propofol and remifentanil were recorded; The resting and motor Visual Analogue Scale (VAS) scores of the patient were recorded at 4, 8, and 24 hours postoperatively. The patient′s serum tumor necrosis factor-α (TNF-α) and melatonin levels were measured on the day before surgery, the evening after surgery, and the first day after surgery. The subjective sleep quality score was recorded on the first day after surgery; The total dosage of sufentanil for patient-controlled intravenous analgesia (PCIA) within 24 hours after surgery, the effective number of compressions (P1) and actual number of compressions (P2) of PCIA, and the number of cases of additional oxycodone analgesia were recorded; The occurrence of respiratory depression, excessive sedation, nausea, and vomiting within 24 hours after surgery was recorded; The occurrence of complications of nerve block in the observation group was recorded. Results:The incidence of delirium during the recovery period in the observation group was lower than that in the control group ( P<0.05). The dosage of propofol and remifentanil in the observation group was lower than that in the control group (all P<0.05), and the awakening time was shorter than that in the control group ( P<0.05). The VAS scores at rest and exercise at 4, 8, and 24 hours after surgery in the observation group were lower than those in the control group (all P<0.05). Compared with the day before surgery, the observation group showed an increase in melatonin levels on the night after surgery ( P<0.05) and an increase in TNF-α levels on the first day after surgery ( P<0.05); The control group had a decrease in melatonin levels on the first day after surgery ( P<0.05), and an increase in TNF-α levels on the night and day after surgery (all P<0.05). The serum melatonin levels in the observation group were higher than those in the control group on the evening and first day after surgery (all P<0.05), and TNF-α levels were lower than those in the control group (all P<0.05). The subjective sleep quality score of the observation group showed that the total dosage of PCIA sufentanil, P1, P2, and the number of cases of additional oxycodone analgesia within 24 hours after surgery were lower than those of the control group (all P<0.05). The incidence of nausea in the control group (13 cases, 38.2%) was higher than that in the observation group (3 cases, 8.8%) ( P<0.05). Conclusions:In elderly patients undergoing unilateral TKA under general anesthesia, preoperative use of ultrasound-guided adductor tube combined with knee joint posterior capsule space block can effectively reduce the occurrence of postoperative delirium.
10.Mechanism of Ginkgo flavone aglycone in alleviating doxorubicin-induced cardiotoxicity based on transcriptomics and proteomics
Yujie TU ; Ying CAI ; Xueyi CHENG ; Jia SUN ; Jie PAN ; Chunhua LIU ; Yongjun LI ; Yong HUANG ; Lin ZHENG ; Yuan LU
China Pharmacy 2024;35(21):2596-2602
OBJECTIVE To investigate the mechanism by which Ginkgo flavone aglycone (GA) reduces the cardiotoxicity of doxorubicin (DOX) based on transcriptomics and proteomics. METHODS Thirty-six mice were randomly assigned to control group (CON group, tail vein injection of equal volume of physiological saline every other day+daily intragastric administration of an equal volume of physiological saline), DOX group (tail vein injection of 3 mg/kg DOX every other day), and GDOX group (daily intragastric administration of 100 mg/kg GA+tail vein injection of 3 mg/kg DOX every other day), with 12 mice in each group. The administration of drugs/physiological saline was continued for 15 days. Mouse heart tissues were collected for RNA-Seq transcriptomic sequencing and 4D-Label-free quantitative proteomic analysis to screen differentially expressed genes and proteins, which were then subjected to Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis. The expression levels of Apelin peptide (Apelin), phosphatidylinositol 3-kinase (PI3K), and protein kinase B (Akt) mRNA and protein in mouse heart tissues, as well as the phosphorylation levels of PI3K and Akt proteins, were verified. H9c2 cardiomyocytes were divided into control group (CON group), DOX group (2 μmol/L), and GDOX group (2 μg/mL GA+2 μmol/L DOX) to determine cell viability and the levels of key glycolytic substances in the cells. RESULTS Six common pathways were identified from transcriptomics and proteomics, including the Apelin signaling pathway, the PI3K-Akt signaling pathway, and insulin resistance. Among them, the Apelin and PI3K-Akt signaling pathways were the most enriched in terms of gene numbers. Target validation experiments showed that compared to the CON group, the relative expression of Apelin, PI3K and Akt mRNA and protein levels, as well as the phosphorylation levels of PI3K and Akt proteins, were significantly decreased in the DOX group (P<0.05 or P<0.01). The relative expression of Apelin, PI3K and Akt mRNA and the phosphorylation levels of PI3K and Akt proteins were significantly increased in the GDOX group as compared with the DOX group (P<0.05 or P<0.01). Cellular experiments indicated that compared to the CON group, cell viability in the DOX group was significantly decreased (P<0.05), the relative uptake of glucose and the relative production of pyruvate and lactate were significantly increased (P<0.05), and the relative production of ATP was significantly reduced (P<0.05). Compared to the DOX group, cell viability in the GDOX group was significantly increased (P< 0.05), and the relative production of pyruvate and lactate was significantly reduced (P<0.05). CONCLUSIONS GA may alleviate DOX-induced cardiotoxicity by upregulating the mRNA and protein expression of Apelin, PI3K, and Akt in heart tissues, and regulating glycolytic processes.


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