1.Expression of COX-2 and p38MAPK of peripheral blood mononuclear cells in trauma patients with multiple organ dysfunction syndrome
Ming WEI ; Ling TU ; Yinghong LIANG ; Jia LIU ; Junhua ZHANG
Chinese Journal of Emergency Medicine 2011;20(6):593-596
Objective To explore the expression of COX-2 and p38MAPK in patients with trauma MODS. Methods Forty MODS patients were evaluated. The levels of peripheral blood mononuclear cells COX-2 and p38MAPK in MODS patients and 40 normal controls was detected by enzyme linked immunosor-bent assay (ELISA). RT-PCR was used to measure the COX-2 mRNA and p38MAPK mRNA expression of in PBMCs. ANOV and correlation analysis were used in statistical analysis. Results The levels of COX-2 and p38MAPK of PBMCs and the mRNA expression in MODS group were higher than in control group (all P <0.05). The levels of COX-2 and p38MAPK of PBMCs and the mRNA expression in dead group were higher than in survival group( all P <0.05). The levels of COX-2 and p38MAPK of PBMCs were positively correlated, (r =0.6 147, P<0.01). The expression of COX-2 mRNA, p38MAPK mRNA of PBMCs and APACHE I scoring were positively correlated (r1 =0.5 009, P1 <0.05,r2 =0. 5 316, P2 <0. 05). Conclusions COX-2 and p38MAPK of PBMCs take part in the onset of MODS, and may service as index to judge the prognosis of MODS.
2.Characteristic analysis of monoblastic sarcoma cutis preceding acute monoblastic leukemia
Zhengjuan ZHAO ; Yinghong YANG ; Wei TIAN ; Yuying WANG ; Lihang LIN
Chinese Journal of Dermatology 2012;45(10):752-753
A 1-year-old boy developed multiple skin-colored nodules on the forehead and extremities when he was 4 months old.Physical examination revealed that his general condition was well with no hepatomegaly,splenomegaly,lymphadenectasis,testicle abnormality or gingival hypertrophy.Pathologically,the epidermis was normal,while the dermis and subcutaneous tissue were diffusely infiltrated with medium-to large-sized deformed cells,which had a small amount of cytoplasm,oval nucleus,irregular shape and fine chromatin.Some infiltrating cells had nuclear groove and nucleoli.Immunohistochemical studies showed that the tumor cells were positive for S-100 protein,CD56,CD123,CD163,CD68,Ki-67 (40%),weakly positive for CD4 (some),but negative for myeloperoxidase,CD1,CD21.Bone marrow smears showed a 24.5% infltration by monoblasts and promonocytes.A diagnosis of monoblastic sarcoma cutis preceding acute monoblastic leukemia was made.
3.Clinical analysis of 272 patients with Kawasald disease
Yinghong ZENG ; Jianping TANG ; Lei SUN ; Ye SHU ; Zhu WEI
Chinese Journal of Dermatology 2009;42(4):234-236
Objective To investigate the clinical features of Kawasaki disease.Methods A retrospective analysis was performed in 272 children diagnosed as Kawasaki disease from 2002 to 2006.Clinical data,laboratory findings and auxiliary examination results were collected for these patients.Results The male-to-female ratio Was 2.58:1.Onset ages between 1 to 3 years accounted for 59.2%of patients.Of these patients,100%had a fever for more than 5 days,76.1%transient polymorphous exanthema,74.6% bilateral conjunctival hyperemia,47.8%flare and fissure on the oral lip,58.5%strawberry tongue,22.8% firm swelling of hands and feet as well as flushing of palms and soles,3 1.2%subacute desquamation at the junctional site between nail bed and skin,36%cervical lymphadenopathy.Laboratory findings showed a significant increase in the count of peripheral blood leukocytes and pefipheral blood platelets as well as erythrocyte sedimentation rate in 80.5%,87.5%and 96.2% of Patients,respectively.Additionally,81.6%of these patients were positive for C reactive protein and the frequency of coronary aaery involvement was 54.3%.All patients were treated with aspirin,and high-dose intravenous immunoglobulin was given to 258 patients.Fever relieved and the condition was controlled in all patients with an average hospitalization period of 8.9 days.Conclusions Kawasaki disease should be suspected in Patients with exanthematous lesions,fever lasting for more than 5 days and poor response to antibiotic therapy.Peripheral blood platelet count and cardiac ultrasound are of great value in the diagnosis of Kawasaki disease.Aspirin iS the first choice in treating Kawasaki disease,and adjunctive high-dose intravennous immunoglobulin treatment may facilitate the quick control offever.
4.Effects of eye acupuncture therapy on neurological function and brain-derived neurotrophic factor expression in a rat model of cerebral ischemia/reperfusion injury
Huiying LIU ; Pengqin WANG ; Ying BIAN ; Jinchun WANG ; Yinghong WEI
Chinese Journal of Tissue Engineering Research 2016;20(18):2634-2641
BACKGROUND: Eye acupuncture therapy is a technique used to adjust qi-blood circulation, relax muscles and tendons, and activate col aterals by acupuncture at the acupoints around the eye bal s and in the orbital border. This therapy has been widely used in the clinic because it exhibits remarkable therapeutic effects on many ischemic cerebrovascular diseases. However, the precise mechanism behind this therapy remains poorly understood. Neurotrophic factors are a protein family including neurotrophic factors and brain-derived neurotrophic factors that can regulate neuronal survival, development and functioning.
OBJECTIVE: To investigate the effects of eye acupuncture therapy on neurological function and nerve growth factor and brain-derived neurotrophic factor expression in the brain tissue of rat models of cerebral ischemia/reperfusion injury.
METHODS: Fifty-four male Sprague-Dawley rats were randomly divided into three groups with 18 rats per group: sham-operated, model and eye acupuncture therapy groups. Rat models of middle cerebral artery occlusion were established by the intraluminal suture method in the model and eye acupuncture therapy groups. Eye acupuncture was performed at the fol owing acupoints liver area, upper-jiao area, lower-jiao area and kidney area located at the internal orbital margin at 2 hours after cerebral ischemia/reperfusion injury. Rat neurological function was evaluated at 3, 7, and 14 days after injury. Nerve growth factor and brain-derived neurotrophic factor expression in the rat brain tissue was detected using immunohistochemical staining method at 1 and 2 weeks after treatment. Cerebral infarct size was determined using TTC staining at 2 weeks after treatment.
RESULTS AND CONCLUSION: (1) At 1 and 2 weeks after injury, nerve growth factor and brain-derived neurotrophic factor expression was significantly greater in the eye acupuncture therapy group than in the model group (P < 0.05), but nerve growth factor and brain-derived neurotrophic factor expression in the eye acupuncture therapy group was decreased at 2 weeks after injury compared to that at 1 week after injury. (2) At 7 and 14 days after treatment, neurological function scores in the eye acupuncture therapy group were significantly lowered, and there was significant difference between eye acupuncture therapy and model groups (P < 0.05), but they were significantly higher than those in the sham-operated group (P< 0.05). (3) At 2 weeks after treatment, cerebral infarct size was significantly greater in the eye acupuncture therapy and model groups than in the sham-operated group (P < 0.01), and it was significantly smal er in the eye acupuncture therapy group than in the model group (P < 0.05). (4)These results indicate that eye acupuncture therapy shows neuroprotective effects on ischemic cerebral injury by increasing nerve growth factor and brain-derived neurotrophic factor expression, improving neurological function, and reducing cerebral infarct size.
5.Function Study of the Nano-Molecular PAMAM as miR Carrier Targeting Gastric Adenocarcinoma
Wei SONG ; Zhongmin JIANG ; Ping XU ; Yinghong LIANG ; Xiaozhi LIU
Tianjin Medical Journal 2013;(12):1147-1149
Objective To study the function of the nano-molecular polyamide-amine (PAMAM) as microRNA(miR) carrier targeting gastric adenocarcinoma, and the foundation of developing an efficient delivery of small molecule drugs tar-geting gastric cancer thereof. Methods The folic acid (FA)/PAMAM comoles compound was prepared by dialysis method. After transfection of miR-7 or liposomes into SGC-7901 cell line, fluorescence microscope was used to detect the gene trans-fect efficiency. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the miR-7 level. The immu-nocytochemistry assay was used to test the protein expressions of epidermal growth factor receptor (EGFR), protein kinase B (PKB) and proliferating cell nuclearantigen (PCNA). The transwell system was utilized to explore the migration ability of tu-mor cells. Results Compared with liposme, FA/PAMAN complex compound can significantly improve the level of miR-7 in SGC-7901 cells,reduce the protein levels of EGFR, PKB and PCNA in SGC-7901 cells, and also reduce the percentage of cancer cell migration (P<0.05). Conclusion PAMAM can effectively transfect miR into gastric cancer cells, which is expected to become an efficient delivery of small molecule drugs.
6.Correlation Study Between Atrial Fibrillation and High-sensitivity Cardiac Troponin T Level in Patients With Stable Coronary Artery Disease
Yinghong TANG ; Qingwei CHEN ; Dazhi KE ; Guiqiong LI ; Wei DENG
Chinese Circulation Journal 2016;31(12):1170-1174
Objective: To explore the relationship between serum level of high-sensitivity cardiac troponin T (hs-cTnT) and atrial ifbrillation (AF) occurrence in patients with stable coronary artery disease (CAD).
Methods: A total of 1011 patients with stable CAD treated in our hospital from 2013-01 to 2015-09 were retrospectively studied. According to quartiles of hs-cTnT, the patients were divided into 4 groups: Group① the patients with hs-cTnT≤7ng/L, n=283, Group② hs-cTnT 7-10ng/L,n=238, Group③ hs-cTnT 10-15ng/L,n=272 and Group④ hs-cTnT>15ng/L,n=218. The relationship between hs-cTnT level and AF occurrence rate was studied; the risk factors for AF occurrence were explored by multi stepwise Logistic regression analysis.
Results: There were 127/1011 (12.6%) patients combining AF and 884 (87.4%) with simple type stable CAD. AF patients had the higher serum level of hs-cTnT than non-AF patients 17.0 (12.0, 25.0) ng/L vs 10.0 (7.0, 13.0) ng/L,P<0.01. With baseline hs-cTnT level increasing, AF occurrence rates were elevated from group① to Group④ as 4.6% (13/283), 4.6% (11/238), 11.0% (30/272) and 33.5% (73/218), Ptrend<0.05. Multi stepwise Logistic regression analysis revealed that with adjusted common risk factors of age and gender, the risk for stable CAD patients suffering from AF in Group④ was 5.324 times higher than group① (95% CI 2.285-12.405,P<0.01).
Conclusion: Increased serum level of hs-cTnT was closely related to AF occurrence in patients with stable CAD.
8.The populations of CD4+CD25highFoxp3+ T regulatory cells, Foxp3 mRNA expression related to disease activity of svstemic Iupus ervthematosus
Ming WEI ; Yaodong ZHANG ; Ling TU ; Yinghong LIANG ; Jia LIU ; Junhua ZHANG
Chinese Journal of Rheumatology 2011;15(11):772-776
ObjectiveTo investigate the ratios of peripheral blood CD4+CD25highFoxp3+ regulatory T cells of systemic lupus erythematosus(SLE) patients,and explore its association with disease activity and nephropathy.MethodsPBMC lymphocytes in 42 patients with SLE and PBMC in 40 normal healthy donors were evaluated for the proportion of Treg cells,as a percentage of the total CD4+ cells,by flow cytometric analysis.Levels of mRNA for Foxp3 were measured with a real-time quantitative PCR.The proportion of Treg cells and its association with SLEDAI,nephropathy,serum anti-dsDNA antibody,and C3 levels were analyzed.Statistical analysis was conducted with t-test and Spearman's correlation analysis.ResultsPatients with active disease had statistically significantly lower levels of CD4+CD25highFoxp3+ Treg than normal controls [(4±3)% vs (7±4)%,P<0.05],while no significant difference could be found between patients with nonactive disease and normal controls(P>0.05).The percentage of peripheral blood CD4+CD25highFoxp3+ Treg/CD4+ in patients with active disease was significantly lower when compared to patients with non-active disease [(9±6)% vs (10±6)%,P<0.05],and it was related to the disease activity.SLE patients with nephropathy had significantly lower levels of CD4+CD25highFoxp3+ Treg and CD4+CD25highFoxp3+ Treg/CD4+ than patients without nephropathy(P<0.05).Foxp3 mRNA levels were lower in PBMC from active disease patients than those in non-active disease.In addition,there was a negative correlation between the populations of CD4+CD25highFoxp3+ and SLEDAI(r=-0.5892,P<0.05).There was a negative correlation between the percentage of CD4+CD25highFoxp3+/CD4+ and SLEDAI (r=-0.4962,P<0.05),while there was a positive correlation between the percentage of CD4+CD25highFoxp3+/CD4+ and C3(r=0.3867,P<0.05).There was a positive correlation between the populations of CD4+CD25highFoxp3+ and Foxp3 mRNA(r=0.6142,P<0.01 ).ConclusionThese suggest that the decrease of CD4+CD25highFoxp3+ Treg and Foxp3 mRNA expression may play a crucial role in the pathogenesis of SLE.
9.Changes of Rho kinase activity in peripheral blood T lymphocytes from patients with atopic dermatitis and their significance
Yinghong LIANG ; Ming WEI ; Ling TU ; Jia LIU ; Yanjie GONG ; Yihua ZHANG
Chinese Journal of Dermatology 2016;49(4):256-260
Objective To evaluate changes of Rho kinase (ROK)activity in peripheral blood T lymphocytes from patients with atopic dermatitis (AD), and to analyze their clinical significance. Methods Eight milliliters of heparin-anticoagulated blood samples were collected from 60 patients with AD and 60 healthy human controls followed by separation of T lymphocytes and sera from these blood samples as well as culture of isolated T lymphocytes with 10% fetal bovine serum for 24 hours. Both patient- and control-derived T lymphocytes were classified into two groups to be cultured with patient- or control-derived sera. In addition, some patient-derived T lymphocytes were classified into 4 groups:Y27632 group treated with the Rho kinase-specific inhibitor Y2763, CD3/CD28 group treated with anti-CD3/anti-CD28 monoclonal antibodies, Y27632 + CD3/CD28 group treated with Y27632 and anti-CD3/anti-CD28 monoclonal antibodies, and control group treated with patient-derived sera. Subsequently, Western-blot analysis was performed to evaluate ROK activity in cells, methyl thiazolyl tetrazolium(MTT)assay to evaluate proliferative activity of T lymphocytes, and ELISA to measure interleukin 6 (IL-6)and IL-10 levels in supernatants of T lymphocytes. Results ROK activity was significantly lower in fresh T lymphocytes from patients than in those from healthy controls (2.47% ± 0.89% vs. 0.65% ± 0.35%, t =2.729, P < 0.05). After 24-hour culture with 10% fetal bovine serum in vitro, ROK activity was significantly decreased in patient-derived T lymphocytes compared with those before culture (0.70% ± 0.38% vs. 2.47% ± 0.89%, t = 2.658, P <0.05), but no significant difference was observed between patient- and control-derived T lymphocytes(0.70% ± 0.38% vs. 0.63% ± 0.32%, t = 1.010, P > 0.05). Compared with T lymphocytes cultured with control-derived sera, those cultured with patient-derived sera showed significantly increased ROK activity (F = 8.22, P < 0.001). Concretely speaking, ROK activity was significantly higher in patient-derived T lymphocytes cultured with patient-derived sera than in those cultured with control-derived sera (2.41% ± 0.87% vs. 0.76% ± 0.41%, P < 0.05), and higher in control-derived T lymphocytes cultured with patient-derived sera than in those cultured with control-derived sera(2.17% ± 0.85% vs. 0.64% ± 0.33%, P< 0.05)at 24 hours. Y27632 could significantly inhibit the proliferation of as well as secretion of IL-6 (F = 18.68, 22.95, respectively, both P < 0.001)by patient-derived T lymphocytes, but had insignificant effects on secretion of IL-10. The cellular proliferative activity and IL-6 supernatant level were significantly lower in the Y27632 group than in the control group, and lower in the Y27632 + CD3/CD28 group than in the CD3/CD28 group (all P < 0.05). Conclusion Aberrant activation of ROK exists in T lymphocytes from patients with AD, which may play a certain role in the pathogenesis of AD.
10.Regulatory effects of miRNA-146a on peripheral blood CD4+ T lymphocytes from patients with psoriasis vulgaris
Ming WEI ; Yinghong LIANG ; Ling TU ; Jia LIU ; Yanjie GONG ; Yihua ZHANG ; Lu YANG
Chinese Journal of Dermatology 2016;49(4):243-247
Objective To evaluate regulatory effects of miRNA-146a on peripheral blood CD4 + T lymphocytes from patients with psoriasis vulgaris, and to investigate the role of miRNA-146a in the pathogenesis of psoriasis vulgaris. Methods Totally, 30 patients with psoriasis vulgaris and 30 healthy human controls were enrolled into this study. Venous blood samples were obtained from these subjects, and CD4 + T lymphocytes were isolated from these samples by using magnetic activated cell sorting (MACS). Real-time quantitative PCR (RT-PCR)was performed to measure the expression of miRNA-146a in peripheral blood CD4+ T lymphocytes, and enzyme-linked immunosorbent assay(ELISA)to determine plasma levels of interferon-γ(IFN-γ)and interleukin 4(IL-4). Some CD4+ T lymphocytes were divided into 3 groups to be transfected with 50 nmol/L negative control miRNA (control group), miRNA-146a mimics(miRNA-146a group)or miRNA-146a inhibitor (miRNA-146a inhibitor group). After 24-hour additional culture, flow cytometry was conducted to determine the number of Th1 and Th2 cells, Western-blot analysis and RT-PCR were performed to measure the protein and mRNA expressions of IFN-γ receptor α (IFN-γRα), T-box expressed in T cells (T-bet)and GATA-binding protein-3 (GATA-3)respectively, and ELISA was carried out to determine the levels of IFN-γ and IL-4 in supernatants of CD4 + T lymphocytes. Results Compared with the healthy control group, the patient group showed significantly increased miRNA-146a expression in peripheral blood CD4 + T lymphocytes (243.81% ± 94.32% vs. 105.74% ± 22.93%, t = 6.653, P < 0.01)and plasma IFN-γ level (27.69 ± 7.64 ng/L vs. 9.75 ± 2.81 ng/L, t = 4.237, P <0.01). Moreover, miRNA-146a expression was positively correlated with plasma IFN-γ level in the patients(r = 0.837, P <0.01). After 24-hour culture in vitro, there was a significant increase in the number of Th1 cells, protein and mRNA expressions of T-bet, and supernatant level of IFN-γ, but a significant decrease in the protein expression of IFN-γRα in the miRNA-146a group compared with the control group (all P < 0.01). However, no significant differences were observed in the number of Th2 cells, mRNA or protein expressions of GATA-3, or supernatant level of IL-4 among the control group,miRNA-146a group and miRNA-146a inhibitor group (all P > 0.05). Conclusion miRNA-146a may play an important role in the pathogenesis of psoriasis vulgaris by participating in the regulation of peripheral blood CD4+ T lymphocytes via affecting Th1 cell differentiation and function.