1.Advances in the study of microRNA in deep vein thrombosis
Journal of Medical Postgraduates 2017;30(8):893-896
Deep vein thrombosis(DVT) refers to a blood clot that forms in a deep vein, and usually occurs in the lower extremities.Typical symptoms include swelling, pain or tenderness in the leg.DVT is one of the common complications of orthopedic surgery and has a complicated pathogenesis.The embolus can fall off into the lung, resulting in the death.MicroRNA (miRNA) is a class of endogenous and non-coding single stranded RNA consisting of 21~23 nucleotides, widely involved in a variety of biological processes.Recent studies have found that microRNA (miRNA) plays a significant role in the pathogenesis of DVT.This article reviews recent advances in the study of miRNAs as novel regulators, diagnostic tools and a therapeutic choice in DVT to provide new insights of its pathogenesis and therapeutic target.
2.Immune regulation of interleukin-23 on peripheral blood natural killer cell in patients with inflammatory bowel disease
Xuan YANG ; Junfen MA ; Zhanju LIU
Chinese Journal of Digestion 2009;29(10):649-652
Objective To study the potential role of interleukin-23(IL-23)in induction of peripheral blood natural killer(NK)cell activation and cytotoxicity in patients with inflammatory bowel disease(IBD),and to investigate its clinical relevance to the pathogenesis.Methods Peripheral blood was collected from 12 healthy subjects,16 patients with Crohn's disease(CD)and 25 patients with ulcerative colitis(UC).NK cells isolated from peripheral blood using immunomagnetie beads were cultured in vitro and stimulated with IL-23 or IgG,or combination of two.Concentrations of tumor necrosis factor α(TNF-α)and interferon-γ(IFN-γ)were determined by enzyme linked immunosorbent assay.NK eytotoxicity to K562 cell was detected by flow cytometry.Results The levels of TNF-α and IFN-γ were not increased in healthy controls under stimulation with IgG or IL-23 (P>0.05),but they were elevated under stimulation with IgG plus IL-23(P<0.05).IL-23,not IgG,was found tO enhance TNF-α and IFN-γ secretion in IBD patients(P<0.05).The levels of TNF-α and IFN-γ were significantly increased in IBD patients under the stimulation of IL-23 plus lgG (P<0.01).The cytotoxicity of NK cells in healthy controls was not observed under stimulation with IL-23 alone or IL-23 plus IgG,whereas it could be enhanced by stimulation with IL-23 in CD patients (62.4%±5.5%)and UC patients(65.3%±6.6%,P<0.05)or with IL-23 plus IgG in CD patients (50.2%±8.7%)and UC patients(51.5%±7.1%,P<0.01).ConclusionIL-23 plays an important role in induction of NK cell activation and cytotoxicity in IBD patients,and may be involved in development of IBD.
3.Relationship between DNA methylation of glucocort icoid receptor gene promotor and glucocorticoid receptor gene mRNA expression in systemic lupus erythematosus patients
Junfen FAN ; Hongbo CHEN ; Hong XIA ; Hongzhen MA ; Xiangfu GAO ; Qiyang SHOU ; Lizong ZHANG ; Yongsheng FAN
Chinese Journal of Rheumatology 2017;21(1):42-45
Objective To investigate the relatio nship between the DNA methylation status of gluco-corticoid receptor (GR) gene promoter and mRNA expression level of GRα gene of peripheral blood mononuclear cells (PBMCs) in patients with systemic lupus erythematosus (SLE). Methods Fifteen new onset SLE patients and fifteen healthy controls were enrolled in this study. The DNA methylation status of GR gene promoter 1 of PBMCs was detected through bisulfite sequencing polymerase chain reaction (PCR). The mRNA expression of GRα, DNA methyltransferases, growth arrest and DNA damage-induced 45α (GADD45α) of PBMCs was detected using the quantitative real-time polymerase chain reaction method. T-test and χ2-test were used to detect the differences between the two groups, Pearson's correlation coefficient was used to analyze the linear correlation between two variables. Results Compared with healthy controls, the mRNA expression of GRα was signi-ficantly declined in SLE patients (10±5, 17±7, t=2.69, P<0.05), and the mRNA expression of DNMT1 and GADD45α was significantly elevated in SLE patients (t=3.11, P<0.05 and t=2.98, P<0.05). The overall mean methylation status of the 142 CpG islands of the four promoters was significantly elevated in SLE patients [(16±8)%vs (11±6)%, t=2.75, P<0.05]. The global methylation status of PBMCs in SLE patients was obviously lower than healthy controls (t=4.39, P<0.05). Conclusion Hypermethylation of GRα promoter may result in GRαgene low expression in PBMCs of patients with SLE.
4.Anticancer effect of mouse marrow-derived dendritic cells transfected by pcDNA3-hCEA
Youtian HUANG ; Ziming DONG ; Mingyao ZHAO ; Hongyan YANG ; Yiguo ZHANG ; Zhimin ZHENG ; Junfen MA ; Liming TANG ; Ning CHEN
Chinese Journal of Pathophysiology 2000;0(11):-
AIM: To prepare the vaccine of DCs(pcDNA3-hCEA) and observ the immunity effect of the DCs(pcDNA3-hCEA) inoculating on CT26(hCEA +) loaded in BALB/c mice. METHODS: DCs were generated from bone marrow in the presence of rmGM-CSF and rmIL-4. A new recombinant plasmid, pcDNA3-hCEA, reformed with inserting a 2.4 kb human CEA cDNA into pcDNA3. DC vaccine was prepared by transfection with pcDNA3-hCEA using lipofectamine. CEA mRNA expressed in DCs(pcDNA3-hCEA) was confirmed by RT-PCR, CEA expression level was detected with RIA method, and CEA specific CTL was induced in vitro . After vaccination of DCs(pcDNA3-hCEA), the survival time of the BALB/c mice challenged with critical loading CT26(hCEA +) was observed. RESULTS: G418 test showed that about 14% DCs were transfected with pcDNA3-hCEA. And CEA mRNA and protein could be detected respectively by RT-PCR and RIA in the genetically modified DCs. Furthermore, the DCs coud be targeted by specific CTL, the survival time of the mice challenged with CT26(hCEA +) was prolonged 1-4 weeks. CONCLUSION: These results demonstrate that specific antitumor immune responses could be induced efficiently by vaccination of DCs(pcDNA3-hCEA), which is transfected eukaryotic expression vector encoding tumor antigen gene.
5.Correlation between serum zinc and brain injury in infants with mild gastroenteritis complicated with benign infantile convulsion and febrile convulsion
Chunmei LI ; Zhenkui LIU ; Yanhui LU ; Wei SHI ; Jiaojiao GAO ; Yanling CAO ; Junfen LIU ; Junshuai MA
Chinese Critical Care Medicine 2023;35(8):860-864
Objective:To investigate the association between serum zinc levels and convulsive brain injury in infants with mild gastroenteritis complicated with benign infantile seizures (BICE) and febrile seizures (FC).Methods:A case-control study method was conducted. 120 children with mild gastroenteritis and convulsion admitted to the First Affiliated Hospital of Hebei North University from January 2020 to January 2022 were enrolled as the research subjects. They were divided into BICE group and FC group according to the type of convulsion. The serum zinc level, the frequency and duration of convulsion, and the occurrence of convulsive brain injury in the two groups were recorded. Multivariate Logistic regression analysis was used to screen the risk factors for convulsive brain injury. The Spearman correlation method was used to analyze the association between serum zinc levels, clinical characteristics of convulsion and convulsive brain injury.Results:A total of 120 children were enrolled, of which 81 developed to BICE and 39 developed to FC during hospitalization. The serum zinc level of children in the FC group was significantly lower than that in the BICE group (μmol/L: 39.24±6.50 vs. 48.65±7.21, P < 0.01). In the BICE group and FC group, the serum zinc level in children with more than 2 convulsions was significantly lower than that in the children with one convulsion (μmol/L: 37.65±6.50 vs. 53.17±7.55 in the BICE group, and 30.27±5.58 vs. 44.16±7.57 in the FC group, both P < 0.01). Serum zinc level in children with convulsion duration ≥5 minutes was significantly lower than that in the children with convulsion duration < 5 minutes (μmol/L: 38.75±6.74 vs. 51.21±7.58 in the BICE group, and 31.08±5.46 vs. 45.19±7.25 in the FC group, both P < 0.01). Moreover, the serum zinc level of children with different convulsion frequency and duration in the FC group was significantly lower than that in the BICE group (all P < 0.01). Among the 120 children, 9 cases of convulsive brain injury occurred, and the incidence rate was 7.50%. The incidence of convulsive brain injury in the BICE group was 1.23% (1/81), which was significantly lower than 20.51% in the FC group (8/39, P < 0.01). The serum zinc level of children with convulsive brain injury was significantly lower than that of children with non-brain injury (μmol/L: 28.50±5.00 vs. 60.22±7.31, P < 0.01), and the number of convulsion was significantly higher than that of non-cerebral injury (≥ 2 convulsions: 100.00% vs. 1.80%, P < 0.01), and the duration of convulsion in children with brain injury was significantly longer than that of non-brain-injured children (convulsion duration ≥5 minutes: 100.00% vs. 11.71%, P < 0.01). Multivariate Logistic regression analysis showed that decreased serum zinc level [odds ratio ( OR) = 2.147, 95% confidence interval (95% CI) was 1.354-3.403], increased number of convulsion ( OR = 3.452, 95% CI was 1.266-9.417), and prolonged convulsion duration ( OR = 3.117, 95% CI was 1.326-7.327) were independent risk factor for convulsive brain injury in children with mild gastroenteritis and convulsion (all P < 0.05). Spearman correlation analysis showed that serum zinc level, convulsion ≥2 times, duration of convulsion ≥5 minutes and convulsion ≥2 times + convulsion duration ≥5 minutes were significantly negatively correlated with the occurrence of convulsive brain injury in FC children ( r values were -0.546, -0.517, -0.522, and -0.528, all P < 0.01). There was no significant correlation between serum zinc level, convulsion ≥2 times, convulsion duration ≥5 minutes and convulsion ≥2 times+convulsion duration ≥5 minutes and convulsive brain injury in BICE children ( r values were -0.281, -0.129, -0.201, -0.243, all P > 0.05). Conclusions:Serum zinc level is related to the characteristics of convulsive symptoms in children with mild gastroenteritis complicated with FC, and has a strong negative correlation with the occurrence of convulsive brain injury. Active targeted intervention and treatment may help reduce the incidence of brain injury in children.