1.Comparative study of transfection of tumor antigen NY-ESO-1 circRNA with a novel cationic lipid-like material C1 to stimulate IFN-γ production in T cells
Hong ZHOU ; Yipeng MA ; Xiaojuan WANG ; Fenglan LIU ; Bin LI ; Dongjuan QIAO ; Xiaojun XIA ; Peigen REN ; Mingjun WANG
Chinese Journal of Microbiology and Immunology 2024;44(9):771-777
Objective:To express NY-ESO-1 epitopes using circular RNA (circRNA) and construct circRNA cancer vaccines using a novel lipid-like material C1, and to evaluate the transfection efficiency and T cell activation potential at cellular level.Methods:In vitro transcription was used to synthesize mRNA and circRNA expressing EGFP and NY-ESO-1 epitopes. Then, they were transfected into COS7 cells and the expression of target proteins were detected in vitro. Lipid-like material C1 and commercial transfection agent TransIT-mRNA were used as delivery systems for mRNA NY-ESO-1 and circRNA NY-ESO-1, and their delivery efficiency was compared. Results:The expression of EGFP was observed under fluorescence microscopy after transfection of mRNA EGFP and circRNA EGFP into COS7 cells for 24 h. The secretion of IFN-γ by T cell receptor-engineered T (TCR-T) cells targeting NY-ESO-1/HLA-A2 was stimulated by COS7-A*02: 01 cells transfected with mRNA NY-ESO-1 and circRNA NY-ESO-1. Compared with mRNA NY-ESO-1, circRNA NY-ESO-1 was able to express the target antigen and stimulate the target cells to release IFN-γ more persistently. The delivery efficiency of C1 material was better than that of commercial transfection reagents when COS7 cells were transfected in vitro. Conclusions:Compared with the linear mRNA, transfection of COS7-A*02: 01 cells with circRNA can lead to more efficient and durable activation of T cells, suggesting that it could be a more suitable candidate for clinical treatment of tumors in the future. The lipid-like material C1 can effectively deliver linear mRNA and circular RNA molecules. This study provides reference for further research on circRNA tumor vaccines.
2.Application value of CT radiomics in differentiating malignant and benign sub-centimeter solid pulmonary nodules
Jianing LIU ; Linlin QI ; Jiaqi CHEN ; Fenglan LI ; Shulei CUI ; Sainan CHENG ; Yawen WANG ; Zhen ZHOU ; Jianwei WANG
Chinese Journal of Radiological Health 2024;33(3):340-345
Objective To investigate the application efficiency and potential of CT radiomics in differentiating malignant and benign sub-centimeter solid pulmonary nodules. Methods A retrospective study was performed on the sub-centimeter ( ≤ 10 mm) solid pulmonary nodules detected by enhanced CT in our hospital from March 2020 to January 2023. Malignancy was confirmed by surgical pathology, and benignity was confirmed by surgical pathology or follow-up. Lesions were manually segmented and radiomic features were extracted. The feature dimension was reduced via feature correlation analysis and least absolute shrinkage and selection operator (LASSO). The 5-fold cross validation was used to validate the model. Support vector machine, logistic regression, linear classification support vector machine, gradient boosting, and random forest models were established for CT radiomics. Receiver operating characteristic curves were drawn. Delong test was used to compare the diagnostic performance of the five classifiers. The optimal model was selected and compared to radiologists with medium and high seniority. Results A total of 303 nodules, 136 of which were malignant, were examined. Radiomics models were established after feature extraction and selection. On test set, the areas under the receiver operating characteristic curves of support vector machine, logistic regression, linear classification support vector machine, random forest, and gradient boosting models were 0.922 (95%CI: 0.893, 0.950), 0.910 (95%CI: 0.878, 0.942), 0.905 (95%CI: 0.872, 0.938), 0.899 (95%CI: 0.865, 0.933), and 0.896 (95%CI: 0.862, 0.930), respectively. Delong test indicated no significant differences in the performance of the five radiomics models, and the support vector machine model showed the highest accuracy and F1 score. The support vector machine model showed significantly higher diagnostic accuracy as compared to radiologists (83.8% vs. 55.4%, P < 0.001). Conclusion The radiomics models achieved high diagnostic efficiency and may help to reduce the uncertainty in diagnosis of malignant and benign sub-centimeter solid nodules by radiologists.
3.Survey on iodine deficiency disorders awareness, education status and willingness of pupils in Guizhou Province
Shuying WANG ; Demei ZHOU ; Chun XIE ; Yang LI ; Fenglan ZHANG ; Li YANG ; Hong XIANG
Chinese Journal of Endemiology 2023;42(8):668-674
Objective:To learn about the awareness, education status and willingness of iodine deficiency disorders (IDD) among elementary school students in Guizhou Province, and to provide a scientific basis for exploring intervention strategies for health education of iodine deficiency in the future.Methods:From June 2021 to May 2022, each IDD monitoring county was selected from the east, south, west, north and middle directions of Guizhou Province, and one elementary school was selected from each county. All students in two classes of Grade 5 and Grade 6 were selected in whole groups to conduct on-site questionnaire surveys in the form of anonymous examinations. The survey mainly included general demographic information and IDD awareness, education status and willingness, and binary logistic regression was used to analyze the relevant influencing factors.Results:A total of 1 259 elementary school students in Guizhou Province were investigated, the rates of awareness of IDD, acceptance of IDD publicity and education, and willingness to accept IDD publicity and education among elementary school students were 37.7% (1 900/5 036), 25.1% (316/1 259) and 69.6% (876/1 259), respectively. By binary logistic regression analysis, gender, residence, grade and father's education level were the influencing factors of pupils' awareness of iodine deficiency ( P < 0.05); residence, age and father's education level were the influential factors of elementary school students receiving iodine deficiency education ( P < 0.05); gender, residence, ethnicity and whether the child was the only child or not were the influential factors of elementary school students' willingness to accept IDD education ( P < 0.05). Conclusions:The elementary school students in Guizhou Province have insufficient knowledge about IDD. The publicity and education for iodine deficiency prevention is limited, and the students' willingness to learn is not high. The publicity, education and intervention for iodine deficiency prevention among elementary school students should be comprehensively strengthened.
4.Epidemic characteristics and influencing factors of influenza-like illness outbreaks in schools in Nanchang City from 2011 to 2022
HE Fenglan ; XIA Wen ; TU Junling ; ZHOU Kun ; LIU Ke ; XIONG Xun ; NI Xiansheng ; ZHOU Xianfeng
China Tropical Medicine 2023;23(11):1146-
Abstract: Objective To analyze the epidemic characteristics and influencing factors of school influenza-like cases in 2011-2022 in Nanchang, so as to provide reference for the prevention and control of school influenza outbreaks. Methods The epidemiological relevant data of school influenza-like cases from 2011 to 2022 in Nanchang and the pathogen test results of respiratory tract samples were collected for epidemiological and etiology analysis. Results From 2011 to 2022, a total of 142 influenza-like cases were reported in schools in Nanchang, with a cumulative morbidity of 2 880 cases and a morbidity rate of 1.89%. A total of 1 263 samples were collected, with an overall positive influenza nucleic acid detection rate of 58.91%. The highest proportion of outbreaks occurred in 2017-2019, while the lowest incidence was in 2011-2013. Outbreaks mainly occurred from November to March of the following year (accounting for 79.58%), presenting obvious seasonal characteristics. The distribution was mainly in primary schools, accounting for 70.42% (100/142) of all outbreaks. From 2011 to 2022, all types of influenza viruses were tested, and more than 2 types of influenza viruses were prevalent each year. The dominant strains alternated between influenza A H1N1, influenza A H3N2 and Victoria lineages of type B influenza viruses. From 2011 to 2019, the influenza epidemic in schools in Nanchang showed a continuous upward trend. During COVID-19 in 2020-2022, Nanchang adopted a variety of non-drug prevention and control measures for COVID-19, and both the number of influenza epidemics in various schools and the number of influenza virus nucleic acid positive cases decreased. The average number of classes involved in the epidemic was (3±2), and the average duration of the epidemic was (8±4) days. Conclusions The outbreaks of influenza-like illness in Nanchang schools is highly prevalent in winter and spring, , with urban primary schools being the high incidence locations. Non-drug prevention and control measures for COVID-19 have an impact on the epidemic trend of influenza, so the continuous monitoring of the school influenza virus activities and improving the timeliness of the report will be conducive to the rapid control of the epidemic.
5.Influence of different doses of clopidogrel combined aspirin on platelet related indexes and safety in UAP patients
Boer AN ; Fenglan ZHOU ; Yafang ZHANG
Chinese Journal of cardiovascular Rehabilitation Medicine 2019;28(2):181-184
Objective :To explore influence of different doses of clopidogrel combined aspirin on platelet related inde—xes and safety in patients with unstable angina pectoris (UAP).Methods : A total of 92 UAP patients treated in our hospital were randomly and equally divided into large dose clopidogrel combined group (received 150mg/d clopi—dogrel combined aspirin ) and routine dose clopidogrel combined group (received 75mg/d clopidogrel combined aspi—rin) ,both groups were continuously treated for three months .Therapeutic effect , medication safety , level of thrombolytic granule membrane protein (GMP)—140 ,maximum platelet aggregation rate (LTA—PAmax ) and ADP—induced maximum platelet amplitude (MAADP ) were compared between two groups .Results : After three—month treatment ,there were no significant difference in total effective rate ,incidence rate of major adverse cardiovascular events ,GMP—140 level ,LTA—PAmax and MAADP between two groups , P>0. 05 all.Incidence rate of severe hemor—rhage in large dose clopidogrel group was higher than that of routine dose group (6.52% vs.2. 17%) without signifi—cant difference ( P=0.609).Conclusion : Increased dose of clopidogrel combined aspirin doesn't obtain significant therapeutic effect ,but may be risk of hemorrhage for unstable angina pectoris ,which calls for clinical attention
6.Improvement Effects of Qingre Jiedu Soft Capsule on Mice Infected by Influenza A H1N1 Virus
Fenglan HE ; Qiang LIU ; Jie ZHOU ; Yuanyuan LIU ; Zhifeng GAO ; Hong XIAO ; Zhanqiu YANG
China Pharmacy 2017;28(4):497-500
OBJECTIVE:To observe the antiviral effects of Qingre jiedu soft capsule(ADSC)against influenza A H1N1 virus in vivo,and to provide a experimental support for clinical therapy of influenza A H1N1 virus. METHODS:BALB/c mice were ran-domly divided into normal control group,model control group,positive drug high-dose,medium-dose and low-dose groups [oselta-mivir phosphate capsule,0.04,0.02,0.01 g/(kg·d)] and ADSC high-dose,medium-dose and low-dose groups [1.5,0.75,0.375 g/(kg·d)].Except for normal control group,others groups were given influenza A H1N1 virus with titer 1.6×10-5.2 via nasal cavity to induce poisoned mice model;6-8 h after modeling,they were given relevant medicine intragastrically,once a day,for 5 days. After medication,the change of body weight within 7 d were observed in mice;the mortality and death prevention rate within 15 d,mean survival days(MSDs)were calculated in mice.Other mice were selected and grouped,and they were given same drugs as above. 8 h after last medication,lung index and inhibition rate of lung index were determined in mice.RESULTS:In model control group,the body weight of mice decreased significantly since 5th day,and mice death was beginning to occur since 8th day(mortal-ity of 85.7% within 15 d);the lung index was increased significantly compared to normal control group (P<0.01). Both ADSC and oseltamivir phosphate capsule could slow down the decrease of body weight in mice,decreased the mortality and lung index of mice,and prolonged MSDs;the MSDs of mice in ADSC high-dose,positive drug high-dose and medium-dose groups were signifi-cantly higher than model control group(P<0.05),and lung index was significantly lower than model control group except that of ADSC low-dose group(P<0.05). CONCLUSIONS:ADSC has certain antiviral effect against influenza A H1N1 virus in vivo.
7.Effects and mechanisms of NECA inhibit endoplasmic reticulum stress to against myocardial ischemia reperfusion injury in rats
Qing WANG ; Yan ZHOU ; Hui HAN ; Fenglan WANG ; Fengmei XING
Journal of Medical Postgraduates 2017;30(6):574-578
Objective Adenosine receptor agonist NECA has a certain myocardial protection, but the specific mechanism is not clear.This paper aimed to study the effect and mechanism of NECA inhibiting endoplasmic reticulum stress to against myocardial ischemia reperfusion injury in rats.Methods 56 Wistar rats of SPF grade were selected and divided into Sham group, I/R group, NECA group and TUDCA group through random number table method.We established the isolated rat heart ischemia reperfusion model by using the Langendorff device.Sham group: heart threaded but not ligated, Kerb-Henseleit buffer continuous infusion 170min;I/R group: heart stability 20min, ischemia 30min, reperfusion 2h;NECA group and TUDCA group: heart stability 20min, ischemia 30min, reperfusion 2h, perfusion solutions containing 0.1μmol/L NECA and 30μmol/L TUDCA were respectively given at 5min before reperfusion, and ended at 30min after reperfusion.Transmission electron microscope was used to evaluate alterations of the myocardial ultrastructures.Western blot analysis was used to detected the expression levels of endoplasmic reticulum stress IREl-XBPl signaling pathway marker protein IRE1α, XBP1s.Immunohistochemical staining was used to detect the expression of IRE1α.Results The results of transmission electron microscopy showed that most of the myofilament ruptured, sarcomere contracture deformation, visible mitochondrial vacuoles degeneration in I/R group, and injury in NECA group and TUDCA group were less than the I/R group, appeared as the filaments arranged more neat, sarcoma only had mild contracture.Immunohistochemical results showed that IRE1αpositive staining was not found in the sham group, and the area of positive staining of IRE1α in I/R group was significantly increased, while the NECA group and TUDCA group were significantly decreased.Compared to the Sham group, the expression level of IRE1α and XBP1s was significantly increased in I/R group(P<0.05);but compared with the expression level of IREα and XBP1s in I/R group(1.72±0.27, 0.97±0.19), the NECA group(1.14±0.16, 0.6±0.13) and the TUDCA(1.07±0.27, 0.58±0.15) group were significantly decreased(P<0.05).Conclusion NECA can reduce endoplasmic reticulum stress through inhibiting IREl-XBPl pathway to protect the myocardium.
8.Prenatal ultrasound assessment for fetal structural abnormalities
Fenglan, WANG ; Jianhua, WANG ; Yunting, ZHANG ; Shihe, KOU ; Jianping, YI ; Mingzhen, DU ; Wei, ZHANG ; Yukun, ZHOU
Chinese Journal of Medical Ultrasound (Electronic Edition) 2015;(6):497-502
Objective To investigate the diagnostic value of prenatal ultrasound for fetal structural abnormalities.Methods The clinical data of 3 101 fetus with structural malformations, diagnosed by prenatal ultrasound and confi rmed after induced labor, and 856 cases of missed diagnosis were analyzed. All these data were collected in Maternal and Child Health Hospital of Tangshan City from January 2010 to June 2014. The types and number of fetal structural abnormalities were recorded. The rates of detection and missed diagnosis for the fetal structural abnormalities were calculated and the reasons for missed diagnosis were analyzed.ResultsA total of 3101 cases (4 171 positions) were diagnosed as fetal structural abnormalities by prenatal ultrasound. The detection rate was 11.51‰ (3 101/269 501). There were 856 cases (947 positions) of fetal structural abnormalities were missed, and the missed diagnosis rate was 3.18‰ (856/269 501). The incidence of fetal structural abmormalities was 14.68‰ (3 957/269 501). In terms of anatomy systems, the top five were cardiovascular system abnormalities (851 cases), the central nervous system abnormalities (691 cases), facial abnormalities (562 cases), urinary system abnormalities (476 cases), and respiratory system abnormalities (134 cases). In terms of single abnormalities, the top five were cleft lip and palate (549 cases), ventricular septal defect (519 cases), hydronephrosis (331 cases), spina bifi da (212 cases), and atrial-ventricular septal defect (163 cases). The top five of missed deformities were polydactylia/symphysodactylia (285 cases), ear deformities (108 cases), hypospadias (93 cases), foot abnormalities (92cases), and ventricular septal defect (52 cases).Conclusions There is a high detection rate for fetal structural abnormalities with prenatal ultrasound. Prenatal ultrasound is an indispensable means for the diagnosis of fetal malformation, but it also has some limitations. There is a high rate of missed diagnosis for polydactylia/symphysodactylia, microtia, and hypospadias.
9.Antiviral activity of Ouyi antipyretic detoxicate soft capsule against influenza a virus H1N1 in vitro.
Fenglan HE ; Qiang LIU ; Yuanyuan LIU ; Jie ZHOU ; Zhifeng GAO ; Hong XIAO ; Zhanqiu YANG
China Journal of Chinese Materia Medica 2011;36(14):1993-1996
OBJECTIVEOur study aims to evaluate the antiviral effects of Ouyi antipyretic detoxicate soft capsule against influenza A virus H1N1 in vivo, so as to find an effective Chinese medicinal formulae for the treatment of the virus infection, which may lay a theoretical foundation for clinic treatment of patient infected with Influenza A Virus H1N1.
METHODWith the observation of cytopathic effect (CPE) that induced by virus ,we investigated viral inhibition rate by MTT colorimetric assay and valued antiviral activity of drugs by therapeutic index (TI) . Meanwhile, Oseltamivir phosphate capsule (Tamiflu) was used as positive control , we carried out experiments through the three ways of preventive effect, direct inactivation and propagation inhibition.
RESULTOuyi antipyretic detoxicate soft capsule could effectively inhibit cytopathic effect (CPE) that induced by Influenza A Virus H1N1. The preventive effect, direct inactivation , and inhibition of endogenous multiplication of Ouyi antipyretic detoxicate soft capsule and Tamiflu against influenza A virus H1N1 were observed. And three types of action therapeutic index (TI) from Ouyi antipyretic detoxicate soft capsule were (15.5 +/- 0.71), (0.55 +/- 0.071), (6.4 +/- 1.27) severally, comparing Tamiflu with (0.4 +/- 0.14), (1.88 +/- 0.29), (4.6 +/- 0.15), respectively.
CONCLUSIONOuyi antipyretic detoxicate soft capsule showed more remarkable preventive effect than Tamiflu in vitro (P<0.01). The possible mechanism of the antiviral activity observed in our study might be the protection of the MDCK cells from viral infection by inhibiting the viral absorption. We need a further study to certify three effects in vivo.
Animals ; Antipyretics ; pharmacology ; Antiviral Agents ; pharmacology ; Capsules ; Cell Line ; Dogs ; Influenza A Virus, H1N1 Subtype ; drug effects ; Inhibitory Concentration 50
10.Application of in situ Y chromosomal hybridization in the detection of mesenchymal stem cells
Fenglan WU ; Xiaofei ZHOU ; Wei CAO ; Chao PENG ; Ailian LIU
Chinese Journal of Tissue Engineering Research 2009;13(45):8924-8928
BACKGROUND:Transplantation of mesenchymal stem cells (MSCs) to multiple sclerosis (MS) is a new developed treatment to be given more and more attention.However,whether the MSCs function by cell replacement after going cross the blood-brain barricr or immune suppression needs further confirmation.OBJECTIVE:To establish a steady and effective method of Y chromosome in situ hybridization (ISH) and to detect the distribution of MSCs in a mouse model with MS.DESIGN,TIME AND SETTING:The observational study was performed at the Laboratory of Molecular Immunology,Institute of Healthy Science from September 2007 to February 2009.MATERIALS:A total of 30 C57BL/6 mice,aged 6-8 weeks and weighing 15-20 g,were selected.Female mice were used to establish experimental autoimmune encephalomyelitis (EAE).METHODS:Y chromosme specific DNA probe labled with digoxine (DIG) was designed and ISH was performed to confirm that the designed probe was Y chromosome specific and biologically sensitive.We transplanted the MSCs from male mice into the female EAE mice,and tracked the MSCs by Y chromosome ISH.MAIN OUTCOME MEASRUES:Distribution of MSCs in mice with EAE was observed under the optical and fluorescence microscope.RESULTS:The probe was confirmed to be Y chromosome specific and biologically sensitive.What's more,a steady and effective method of ISH was established.Some MSCs were detected in the periphery organs-spleen and lymph node of mice with EAE;seldom in spinal cord which indicated that MSCs might play its roles by immune suppression.CONCLUSION:Some MSCs were detected in the periphery organs-spleen and lymph node of mice with EAE,but seldom in spinal cord in the central nervous system.

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