1.A new variant of respiratory syncytial virus glycoprotein gene and clinical features of infected children
Mingli JIANG ; Fengjie WANG ; Zhenzhi HAN ; Yanpeng XU ; Yu SUN ; Runan ZHU ; Dongmei CHEN ; Qi GUO ; Yutong ZHOU ; Linqing ZHAO
Chinese Journal of Microbiology and Immunology 2024;44(2):120-127
Objective:To analyze the epidemiological features of respiratory syncytial virus (RSV) infection in Beijing, and monitor the sequence variations in RSV glycoprotein (G) gene and clinical features of infected children.Methods:Respiratory tract specimens were collected from children with acute respiratory infection in the Children′s Hospital Affiliated to Capital Institute of Pediatrics from January 1, 2023 to December 31, 2023. RSV-positive specimens screened by multiple nucleic acid testing were subjected to PCR to amplify the full-length RSV G gene. A phylogenetic tree was constructed after gene sequencing to analyze RSV subtypes and trace G gene variants. Clinical data were retrieved from the medical record system to analyze the clinical features of children with RSV infection in Beijing.Results:A total of 5 489 respiratory specimens were collected from 3 046 male patients and 2 443 female patients. The average age of the patients was 4.36 years. A total of 589 RSV-positive specimens (10.7%, 589/5 489) were detected with 349 from male patients and 240 from female patients. The average age of children with RSV infection was (2.51±2.78) years and the median age was 0.48 years. RSV had been circulating among children in Beijing since March 2023 with two epidemic peaks in May (24.6%, 122/496) and December (18.2%, 126/693). The predominant subtype of RSV in the first half of 2023 was subtype A, but it was replaced by subtype B from November 2023. Phylogenetic analysis revealed a novel G gene of RSV subtype B (RSV-B-BA9-954bp) with a length of 954 bp, which belonged to a new cluster in the phylogenetic tree. The percentage of patients admitted to the Intensive Care Unit (ICU) was higher in children with new variant of RSV subtype B infection than in those with common RSV subtype B infection [44.1% (15/34) vs 25.2% (31/123), χ 2=4.600, P=0.032], while the counts of white blood cells and the levels of C-reactive protein were lower in the children with new variant infection ( P<0.05). Conclusions:RSV has been prevalent among children in Beijing since March 2023 with two epidemic peaks. The predominant A subtype is gradually replaced by to B subtype. A new variant of RSV B G gene (RSV-B-BA9-954bp) is detected among the children.
2.Transcriptomic analysis of A549 cells infected with ON1 genotype of human respiratory syncytial virus subtype A isolated in Beijing
Fengjie WANG ; Mingli JIANG ; Qi GUO ; Xiaoyun LI ; Yu SUN ; Runan ZHU ; Linqing ZHAO
Chinese Journal of Microbiology and Immunology 2024;44(2):128-133
Objective:To analyze the differentially expressed genes of human respiratory syncytial virus (RSV) subtype A genotype ON1, a predominant genotype in Beijing, after infecting A549 cells using transcriptomic sequencing, and provide potential targets for RSV prevention and treatment.Methods:A local strain (61397-ON1) identified by whole-genome sequencing as ON1 genotype of RSV subtype A was selected to infect A549 cells. Total mRNA was extracted, and the differentially expressed genes in infected and uninfected A549 cells were screened by transcriptomic sequencing. GO analysis and KEGG pathway analysis were performed. Besides, six genes with differential expression ratio greater than two times were randomly selected for qRT-PCR verification.Results:There were 1 632 differentially expressed genes between infected and uninfected A549 cells, of which 807 genes were up-regulated and 825 genes were down-regulated. The differentially expressed genes were mainly involved in immune response-related biological processes such as cytokine response and positive regulation of MAPK cascades, and were enriched in MAPK signaling pathway, NOD-like receptor signaling pathway, p53 signaling pathway, TNF signaling pathway, IL-17 signaling pathway and NF-κB signaling pathway. The results of qRT-PCR for six differentially expressed genes were consistent with the trend of transcriptome data.Conclusions:The differentially expressed genes of RSV A subtype ON1 genotype after infecting A549 cells are mainly involved in cytokine response and immune-related signaling pathways. This study provides basic data for further study of the molecular mechanism of RSV infection and the development of prevention and treatment strategies.
3.Significance of lysophosphatidic acid receptor 6 in the large-cell transformation of mycosis fungoides and its effect on the proliferation and apoptosis of cutaneous T-cell lymphoma cells
Yuchieh LIN ; Fengjie LIU ; Yumei GAO ; Xiangjun LIU ; Bufang XU ; Yingyi LI ; Pan LAI ; Zhuojing CHEN ; Jingru SUN ; Ping TU ; Yang WANG
Chinese Journal of Dermatology 2022;55(2):102-109
Objective:To determine lysophosphatidic acid receptor 6 (LPAR6) expression in patients with mycosis fungoides (MF) , a variant of cutaneous T-cell lymphoma (CTCL) , and to investigate its role and mechanism of action in the development and prognosis of CTCL.Methods:A total of 110 patients with confirmed MF were collected from Department of Dermatology, Peking University First Hospital from 2011 to 2020, including 24 with large-cell transformation (LCT) and 25 with non-large cell transformation (NLCT) in the discovery cohort, and 24 with LCT and 37 with NLCT in the validation cohort. RNA sequencing and RT-PCR were conducted to determine the LPAR6 expression in patients in the discovery cohort and validation cohort respectively. LPAR6 expression was compared between patients with LCT and those with NLCT, and its effect on the prognosis of patients was evaluated. Two LPAR6-overexpressing CTCL cell lines MyLa and Sz4 were constructed to evaluate the effect of LPAR6 overexpression on proliferative activity of MyLa and Sz4 cells, with the cells normally expressing LPAR6 as the control group; after the treatment with LPAR6-related ligand lysophosphatidic acid (LPA) , 2S-OMPT, adenosine triphosphate (ATP) or adenosine (ADO) , the effects of LPAR6 activation on the proliferative activity and apoptosis of LPAR6-overexpressing MyLa and Sz4 cells were evaluated by the MTS method and flow cytometry respectively. Log-rank test was used for prognostic analysis, and t test or Mann-Whitney U test was used for comparisons between two groups. Results:As RNA sequencing showed, LPAR6 was one of the significantly underexpressed genes in the LCT group in the discovery cohort; in the validation cohort, LPAR6 expression (median[ Q1, Q3]) was significantly lower in the LCT group (204.90[81.90, 512.70]) than in the NLCT group (809.40[417.50, 1 829.20], U= 242.00, P= 0.002) ; in the two cohorts, the underexpression of LPAR6 was significantly associated with increased risk of poor prognosis (both P < 0.01) . Cell proliferation assay showed no significant difference in the proliferative activity of MyLa or Sz4 cells between the LPAR6 overexpression group and control group at 0, 24, 48 and 72 hours during the experiment (all P > 0.05) ; 48 hours after activation of LPAR6 by LPA, 2S-OMPT, ATP and ADO in MyLa cells, the LPAR6 overexpression group showed significantly decreased cellular proliferative activity (1.38 ± 0.01, 1.04 ± 0.01, 1.09 ± 0.03, 1.23 ± 0.01, respectively) compared the control group (1.73 ± 0.04, 1.23 ± 0.01, 1.24 ± 0.01, 1.42 ± 0.03, t= 30.33, 18.38, 4.78, 5.75, respectively, all P < 0.05) , but significantly increased cell apoptosis rate (17.93% ± 0.88%, 17.75% ± 0.35%, 23.97% ± 0.57%, 31.44% ± 0.34%, respectively) compared the control group (3.98% ± 0.03%, 7.81% ± 0.59%, 11.95% ± 0.85%, 12.02% ± 0.48%, t= 15.93, 14.49, 11.74, 33.01, respectively, all P < 0.05) ; 48 hours after activation of LPAR6 by 2S-OMPT and ADO in Sz4 cells, compared with the control group, the LPAR6 overexpression group also showed significantly decreased cellular proliferative activity (2S-OMPT: 1.29 ± 0.04 vs. 1.48 ± 0.01; ADO: 1.27 ± 0.01 vs. 1.51 ± 0.02; both P < 0.05) , but significantly increased cell apoptosis rate (2S-OMPT: 41.70% ± 0.70% vs. 29.35% ± 0.55%; ADO: 37.05% ± 0.15% vs. 24.60% ± 1.00%; both P < 0.05) . Conclusions:LPAR6 was underexpressed in the patients with LCT, and its underexpression was significantly associated with increased risk of poor prognosis. In vitro activation of LPAR6 could inhibit the proliferation of CTCL cells and promote their apoptosis, suggesting that the decrease of LPAR6 expression may be one of the important mechanisms underlying disease progression in patients with LCT.
4.Mechanism of improving skin barrier function in a mouse model of atopic dermatitis using Mahuang Lianqiao Chixiaodou decoction and its disassembled formula
Liu JINGANG ; Sun YAN ; Yuan HUIMIN ; Li YUDA ; Feng JING ; Gao YUSHAN ; Tang YANG ; Zhang SHUJING ; Wang HANG ; Zheng FENGJIE
Journal of Traditional Chinese Medical Sciences 2021;8(4):309-316
Objective:To reveal the mechanism of Mahuang Lianqiao Chixiaodou decoction and its disassembled formula for improving the skin barrier function in a mouse model of atopic dermatitis (AD).Methods:Sixty specific-pathogen free male BALB/c mice were randomly divided into the control group,model group,whole formula group (WF),exterior-releasing formula group (ERF),interior-clearing for-mula group (ICF),and positive control group (PC).A mouse model of AD was established using the semi-antigen 2,4-dinitrofluorobenzene induction method.The lesion scores,transepidermal water loss and pH,and skin histopathology of mice in each group were observed.The expressions of filaggrin,loricrin,and involucrin were detected by the streptavidin peroxidase immunohistochemical method and western blotting,and their mRNA expressions were detected by quantitative polymerase chain reaction.Results:Mice in the WF,ERF,ICF,and PC groups showed reduced skin lesion performance,improved histopathology,decreased skin lesion score,transepidermal water loss and pH,and upregulated ex-pressions of proteins including filaggrin,loricrin,and involucrin,and their mRNAs.The most obvious regulatory effect was observed in the WF group,followed by the ICF,ERF,and PC groups,accordingly.Conclusions:Mahuang Lianqiao Chixiaodou decoction and its disassembled formula can improve the skin barrier function in a mouse model of AD by upregulating filaggrin,loricrin,and involucrin,and their mRNA expressions,and the most optimal effect was noted in the WF group,followed by the ICF and ERF groups,which suggests that the effect of clearing heat and resolving dampness in improving the skin barrier function of AD is more obvious and is one of the key treatments for AD.
5.Clinical features and outcome of posttraumatic endophthalmitis in children
Xiaozhen HE ; Fengjie LI ; Peiyan SHI ; Jijun SUN ; Weiyun SHI ; Ting WANG
Chinese Journal of Experimental Ophthalmology 2019;37(7):553-558
Objective To investigate the clinical feature and treatment outcome of paediatric posttraumatic endophthalmitis.Methods A retrospective case study was performed.Twenty-six paediatric posttraumatic endophthalmitis children with 26 eyes were enrolled in Shandong Eye Hospital from April 2014 to April 2017.The clinical features,causes of trauma,time and mode of treatment,complications and visual acuity were analyzed.The subjects were graded according to the prognostic status,and the prognosis of the children was compared among different treatment time,vitrectomy time,operation frequency and antibiotic application.Results Of 26 cases,including 20 males and 6 females,aged from 1 year to 13 years,the average age was (6.56±3.25) years.The subjects were divided into infant group (2 cases,7.69%),preschool group (10 cases,38.46%) and school age group (14 cases,53.85%).The main cause of injury was wire,accounting for 34.62% (9/26).Syringe needles accounted for 15.38% (4/26).Fireworks accounted for 11.54% (3/26).All of the 26 cases received vitrectomy,24 cases combined with lens extraction (92.31%).Among the 26 cases,8 cases had no primary visual acuity record;in the remaining 18 cases,the visual acuity at last follow-up was significantly improved when compared with the preoperative primary visual acuity (x2 =7.385,P =0.033).The positive rate of pathogenic bacteria culture in aqueous humor and vitreous humor was 23.08% (6/26),gram-positive cocci was the main pathogenic bacteria of posttraumatic endophthalmitis,which account for 83.3% (5/6).The prognostic grade was grade 1 in 15 cases (57.7%),grade 2 in 7 cases (26.9%) and grade 3 in 4 cases (15.4%).There were significant differences in the number of prognostic grading cases among children with different hospitalizing time or vitrectomy time (all at P<0.05).The hospitalizing time,time of vitrectomy,number of operations and systemic application of antibiotics were positively correlated with the prognosis grade (rs =0.580,0.414,0.428,0.535;all at P < 0.05).Conclusions Children with posttraumatic endophthalmitis should undergo vitrectomy as early as possible,which can receive better prognosis.
7.Correlation analysis of methylation status of UNC5C gene promoter with colorectal cancer UNC5C
Dongsheng SUN ; Chunyan GAO ; Zhigang WANG ; Ning LIU ; Fengjie JIA
Chongqing Medicine 2015;(11):1487-1489
Objective To investigate the correlation between methylation status of UNC 5C gene promoter with colorectal cancer UNC5C .Methods 54 cases of sporadic colorectal cancer and related normal mucosal tissue ,as well as 6 colon cancer cell lines and fiber cell lines(FCL) in the oncology department of the Kailuan General Hospital from February 2010 to March 2013 were selected as the research objects and performed the mRNA and methylation analysis for exploring the correlation between the netrin‐1 receptor UNC5C defects with colorectal cancer disease .Results mRNA of UNC5A and UNC5B was expressed in the detected colorectal cancer cell lines .Except FCL for UNC5C ,all of the cancer cell lines had no mRNA expression .In colorectal cancer tissue , UNC5C methylation was significantly higher than that in the normal mucosa ,the difference was statistically significant (P<0 .05) . The methylation levels of UNC5C gene in the stage Ⅰ to Ⅱ ,and the stage Ⅲ to Ⅳwere significantly higher than non‐methylation levels ,the differences were statistically significant (P<0 .05) .The correlation analysis by the Spearman method showed that the UNC5C defects was positively correlated with colorectal cancer disease (r= 0 .856 ,P< 0 .05) .Conclusion Netrin‐1 receptor UNC5C defect has certain correlation with colorectal cancer disease .
8.Expression of transcriptional factor Hand2 in fetal rats with gestational diabetes mellitus during cardiac development
Xia WU ; Wanyi HUANG ; Shasha HAN ; Fengjie SUN ; Jing XU ; Xuesong YANG ; Guosheng LIU
Chinese Journal of Pathophysiology 2014;(10):1765-1771
AIM:To study the effect of Hand2 (one of basic helix-loop-helix proteins’ transcription factors) expression on the development of the cardiac tissues in the fetal rats from gestational diabetes mellitus ( GDM) parents, and to investigate the potential pathogenesis of GDM-induced congenital cardiac defects in rats.METHODS: The adult Spra-gue-Dawley female rats were randomly divided into blank control group (n=24), GDM group (n=30), negative control group ( n=30) and insulin intervened group ( n=30) .The GDM model was established by intraperitoneal injection of 2%streptozotocin (STZ, 40 mg/kg body weight) to the pregnant rats on the successive day.The rats in insulin intervened group were injected with intermediate-acting insulin in order to keep the fasting blood glucose in the normal range.The rats in negative control group were injected with citric acid-sodium citrate buffer solution in the same position.Blood glucose and body weight were examined every day 72 h after STZ injection.On E12, E15 and E19, the rats were anesthetized and the embryonic cardiac tissues were collected after caesarean section.The histopathological changes of the cardiac tissues were observed under microscope with HE staining.The expression of Hand2 was analyzed by the method of immunohisto-chemistry.The expression of Hand2 in the cardiac tissues at mRNA and protein levels was determined by real-time fluores-cence quantitative PCR and Western blotting.RESULTS:During the development of embryonic heart, the protein expres-sion of Hand2 in the cardiac tissues was showed dynamic changes.Observed on E12, obviously increased on E15, and at the highest level on E19.Compared with the other 3 groups, the protein and mRNA expression of Hand2 in GDM group was decreased at the time points of E12 and E15.CONCLUSION:The morbidity of fetal cardiac malformation is significantly increased in GDM group, suggesting that Hand2 may be involved in the development of cardiac malformation in GDM.
9.Comparison of HBsAg Results between Roche Cobas e601 and Abbott Architect i2000 analysis systems
Bin SUN ; Kang LI ; Chun WU ; Fengjie GUO ; Guangliang BAI ; Mei DONG
International Journal of Laboratory Medicine 2014;(20):2823-2824,2851
Objective To compare the difference of the HBsAg detected results between the Roche cobas e601 electrochemilumi-nescence immunoassay instrument and the Abbott Architect i2000chemiluminescent microparticle immunoassay instrument.Methods The HBsAg positive specimens with the quantitation results of lower than 250IU/mL detected by the Abbott Architect i2000 were selected and simultaneously detected by the Roche cobas e601.The differences of detected results were compared and per-formed the linear correlation and analysis regression.Results 46 clinical specimens were detected.The detected results had best correlation between the two instruments by getting rid of 1 specimen with unconformable reactivity of detected results.15 speci-mens had the HBsAg detected result of 0.05-1.00 IU/mL by the Abbott Architect i2000,the linear regression equation was Y =17.49X+0.843(r=0.979);15 specimens had the HBsAg detected result of 1.1 -10.00 IU/mL IU/mL by the Abbott Architect i2000,the linear regression equation was Y =15.72X +21.06(r=0.952);15 specimens had the HBsAg detected result of 11 -250 IU/mL by the Abbott Architect i2000,the linear regression equation was Y =29.17X -129(r=1.000).Conclusion The detected results have better correlation between the two instruments and can be mutually converted by the formulas.
10.Clinical application of poly-L-lactide sternal coaptation pins in coronary artery bypass grafting
Fengjie LI ; Weiyan SUN ; Feng XIAO
Chinese Journal of Tissue Engineering Research 2013;(47):8288-8293
BACKGROUND:Poly-L-lactide sternal coaptation pins have been gradual y used in clinic, but there are stil few reports about the clinical effects of poly-L-lactide sternal coaptation pins in sternal fractures and sternal fixation after heart surgery.
OBJECTIVE:To summarize the clinical effects of poly-L-lactide sternal coaptation pins in coronary artery bypass grafting.
METHODS:Total y 150 patients who had received coronary artery bypass graft were enrol ed. Ninety-five out of 150 patients received sternal fixation using steel wires as control group, and the other 55 patients underwent sternal fixation using steel wires and poly-L-lactide sternal coaptation pins as test group. Pethidine dosage, incidence rates of pulmonary complications and wound infection, postoperative hospitalization days and hospital costs were compared between the two groups.
RESULTS AND CONCLUSION:During the fol ow-up period (3-8 years), there was no sternal infection and nonunion in the two groups. In the test group, pethidine dosage and hospitalization days were fewer than those in the control group (P<0.05). No significant differences were found in the incidence rates of complications and wound infection and hospital costs between the two groups (P>0.05). These findings indicate that poly-L-lactide sternal coaptation pins are feasible in the coronary artery bypass grafting.

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