1.Study on the expression of peripheral blood IL-23/Th17 axis in patients with Psoriasis Vulgaris treated with Narrow band Ultraviolet B
Xiaolan GUO ; Xiujuan LI ; Wenbiao YANG ; Fangyi HUANG ; Zhibin OUYANG
Journal of Chinese Physician 2012;14(6):736-738,741
ObjectiveTo investigate the therapeutic effect of narrow band ultraviolet B ( NB VUB)and its influence on the expression of IL23/Th17 axis in peripheral blood of patients with Psoriasis vulgaris,and to further explore the mechanism of action of NB UVB.MethodsForty - eight cases of Psoriasis vulgaris were treated with NB-UVB irradiation for 20 times,the therapeutic effect was evaluated by the Psoriasis Area and Severity Index (PASI) scores.Peripheral blood was obtained from normal healthy controls and patients with Psoriasis vulgaris before and after NB-UVB irradiation.Three color flow cytometry was carried out to quantify Thl7 cells,and ELISA was used to examine the levels of serum IL17 and IL23.Results The mean PASI scores were significantly decreased after treatment with NB UVB irradiation[ (8.12±4.05)score vs (3.98±2.03) score,P<0.01 ].Levels of Th17[ (2.78 ± 1.93)% vs (0.98±0.56)% ],IL-17[ (23.85±7.98) pg/ml vs (6.53±4.26) pg/ml] and IL-23 [ (29.73 ± 12.08)pg/ml vs ( 16.73±8.91 )pg/ml] were significantly higher in patients with Psoriasis vulgaris than that in healthy controls ( P <0.01 ).After treatment with NB-UVB irradiation,levels of Th17 [ ( 1.13 ± 0.51 ) % ],IL-17 [ ( 8.03±5.01 )pg/ml ],and IL-23 [ ( 17.03 ± 9.85 )pg/ml ] were significantly decreased than before ( P < 0.01 ),and were positively correlated with PASI ( P <0.05).ConclusionsNB-UVB may affect IL23/IL17 to achieve its therapeutic effect on patients with Psoriasis vulgaris.
2.Effects of lincRNA-cox2 on the polarization of murine RAW264. 7 macrophages
Zikun HUANG ; Fangyi YAO ; Qing LUO ; Jianqing YE ; Zhen DENG ; Yang GUO ; Hong JIANG ; Junming LI
Chinese Journal of Microbiology and Immunology 2016;36(12):881-886
Objective To investigate the effects of lincRNA-cox2 on the polarization of murine RAW264. 7 macrophages by analyzing the expression of lincRNA-cox2 in RAW264. 7 macrophages of M1 and M2 phenotypes. Methods Murine RAW264. 7 cells were induced by IFN-γand LPS to polarize to M1 phenotype, and were induced by IL-4 to polarize to M2 phenotype. The expression of lincRNA-cox2 in M1 and M2 macrophages were analyzed by real-time quantitative PCR ( RT-PCR) . We designed and synthesized siRNA oligo for lincRNA-cox2 and unrelated sequences. Then the siRNA oligo and NC oligo were transfected into RAW264. 7 cells by LipofectmineTM 2000. The transfected RAW264. 7 cells were induced by IFN-γand LPS or by IL-4 to polarize to M1 or M2 macrophages. Enzyme linked immunosorbent assay ( ELISA) was performed to measure the secretion of IL-10 and IL-12 induced in different conditions. The expression of in-ducible nitric oxide synthase ( iNOS ) , TNF-α, arginase 1 ( Arg-1 ) and found in inflammatory zone 1 (Fizz1) at mRNA level were detected by RT-PCR. The M1 macrophages were transfected with siRNAs to knock down the expression of lincRNA-cox2 for analyzing the biological effects of lincRNA-cox2 on the polar-ization of macrophages. Results The relative expression of lincRNA-cox2 in M1 macrophages was signifi-cantly higher than that in RAW264. 7 cells and M2 macrophages. Compared with the control group, the RAW264. 7 cells transfected with lincRNA-cox2-siRNA showed decreased secretion of IL-12 and inhibited expression of iNOS and TNF-αat mRNA level after IFN-γand LPS induction, but increased secretion of IL-10 and enhanced expression of Arg1 and Fizz1 at mRNA level after IL-4 induction. Transfecting the M1 mac-rophages with lincRNA-cox2-siRNA inhibited the secretion of IL-12, but promoted the secretion of IL-10. Conclusion This study indicated that lincRNA-cox2 was involved in the regulation of macrophage pheno-types by promoting the polarization to M1 macrophages and inhibiting the polarization to M2 macrophages.
3.Differential expression profile of long non-coding RNA in the lipopolysaecharide-induced inflammation of monocyte-derived macrophages
Zhen DENG ; Fangyi YAO ; Jianqing YE ; Jianqing XU ; Cheng QING ; Qing LUO ; Zikun HUANG
Chinese Critical Care Medicine 2017;29(4):306-310
Objective To analyze the expression profile of long non-coding RNA (lncRNA) in the lipopolysaecharide (LPS)-induced inflammation of monocyte-derived macrophages.Methods Peripheral blood mononuclear cells were derived from healthy donor and induced into macrophages. The macrophages were divided into blank control group and LPS (1 mg/L) stimulated 12 hours group. Culture supernatants and cell pellets were harvested in each group, enzyme linked immunosorbent assay (ELISA) was used to assay the production changes of interleukins (IL-1β and IL-6), and tumor necrosis factor-α (TNF-α) in the supernatant. The technique of lncRNA microarray was used to test the lncRNA expression profile in LPS-induced inflammation of macrophages and control macrophages. The raw data of lncRNA were pretreated for normalization. Five lncRNA expressions were validated by real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Furthermore, qRT-PCR was used to detect the expression of NR_028034 in macrophages after LPS-induced inflammation.Results ① The contents of IL-1β (ng/L:562.93±61.17 vs. 59.74±15.68), IL-6 (ng/L: 702.46±92.31 vs. 71.66±18.25) and TNF-α (ng/L: 794.50±63.89 vs. 85.12±22.07) in the LPS group were significantly higher than those in the blank control group (allP < 0.01). These results indicated that the inflammatory model of human macrophages was constructed successfully. ② Compared with blank control group, and 1479 lncRNA which have more than 2 folds variation and significant difference (P < 0.05) by statistical analysis was defined as lncRNA with differential expression. Among these lncRNA, LPS group showed 953 up- regulated and 526 down- regulated genes by 2 folds and 49 up- regulated and 35 down- regulated genes by 5 folds. ③ qRT-PCR results were generally consistent with the microarray data. ④ The expression of NR_028034 was increased by (4.41±0.65), (11.56±2.04), (18.58±1.36) folds compared with blank control group at 3, 6, 12 hours after LPS stimulation (allP < 0.01).Conclusions These data show a significantly altered lncRNA expression profile in the LPS-induced inflammation of monocyte-derived macrophages, suggesting that lncRNA may be involved in regulation of macrophages inflammatory response.
4.Clinical research of autohemotherapy combined with bacillus calmette-guerin polysaccharide and nucleic acid in treatment of facial steroid-dependent dermatitis
Zudong MENG ; Fangyi WU ; Yahong WANG ; Xia HUANG ; Yu WANG ; Xiaolan WANG ; Wanxiang SHENG
Chinese Journal of Postgraduates of Medicine 2013;36(30):7-8
Objective To observe the clinical effect and security of autohemotherapy combined with bacillus calmette-guerin polysaccharide and nucleic acid in treatment of facial steroid-dependent dermatitis.Methods The 74 patients with facial steroid-dependent dermatitis were divided into treatment group (38patients) and control group (36 patients) according to the treatment method.The patients in treatment group were treated with autohemotherapy combined with bacillus calmette-gnerin polysaccharide and nucleic acid 1 ml muscle injection,every 3 days a time for 4 weeks.The patients in control group were treated with bacillus calmette-guerin polysaccharide and nucleic acid,every 3 days a time for 4 weeks.The effect was compared between the two groups.Results The total effective rate in treatment group was 84.2%(32/38),in control group was 63.9%(23/36),there was significant difference between the two groups (P < 0.05).No serious adverse reaction was found in two groups.Conclusion Autohemotherapy combined with bacillus calmettegnerin polysaccharide and nucleic acid is safe and effective in treatment of facial steroid-dependent dermatitis.
5.Expression profile of circular RNA in LPS-treated murine peritoneal macrophages
Qing LUO ; Fangyi YAO ; Zhen DENG ; Rigu SU ; Lan LIAO ; Zikun HUANG
Chinese Journal of Microbiology and Immunology 2017;37(6):405-411
Objective To analyze the expression profile of circular RNA (circRNA) in LPS-treated murine peritoneal macrophages (MPMs) and to investigate the effects of mmu_circ_0000790 (circ790) on the secretion of pro-inflammatory cytokines in MPMs following LPS stimulation.Methods MPMs were isolated from C57BL/6 male mice and then stimulated with (LPS treatment group) or without (blank control group) 1 mg/L of LPS for 12 hours.Supernatants of cell culture and cell pellets were collected from each group.Enzyme linked immunosorbent assay (ELISA) was used to measure the changes in the secretion of IL-1β,IL-6 and TNF-α in the supernatants.Expression profile of circRNA in macrophages from the two groups (n=3) was analyze by circRNA microarray.Some differentially expressed circRNAs were validated by real-time PCR (RT-PCR).Moreover,RT-PCR was also performed to detect the expression of circ790 in MPMs after LPS stimulation.Small interfering RNA (siRNA) oligo specific for circ790 was designed and synthesized.Then the synthesized siRNA oligo and normal control (NC) oligo were transfected into MPMs by LipofectamineTM2000.The transfected MPMs were treated with LPS.ELISA was used to detect the levels of IL-1β,IL-6 and TNF-α in the supernatants.Arraystar's home-made microRNA target prediction software was used to predict circ790/microRNA.Results The concentrations of IL-1β,IL-6 and TNF-α in the LPS treatment group were significantly higher than those in the blank control group (P<0.01).These results indicated that the inflammatory model of MPMs was successfully constructed.Statistical differences in 34 differentially expressed circRNAs were found between the two groups (P<0.05).Among them,12 circRNAs were up-regulated and the other 22 circRNAs were down-regulated in the LPS treatment group.RT-PCR results were generally consistent with the microarray data.The expression of circ790 was increased by (1.94±0.15),(3.18±0.13) and (4.21±0.22) folds as compared with that of the blank control group at 3,6 and 12 h after LPS stimulation (P<0.05).After transfecting the MPMs with circ790-siRNA,the secretion of IL-6 was significantly decreased (P<0.05) without notable influence on the secretion of IL-1β and TNF-α.circ790 could function as a microRNA sponge to regulate the gene expression.Conclusion These data show a significantly altered circRNA expression profile in the LPS-treated MPMs.circ790 may be involved in the regulation of IL-6 secreted by macrophages.
6.Clinical value of serum long non-coding RNA metastasis-associated lung adenocarcinoma transcript 1 in the diagnosis of pulmonary tuberculosis
Qing LUO ; Fangyi YAO ; Yiping PENG ; Rigu SU ; Zhen DENG ; Zikun HUANG
Chinese Journal of Infectious Diseases 2017;35(11):684-687
Objective To detect the serum level of long non-coding RNA (lncRNA ) metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) in pulmonary tuberculosis (TB) patients ,and to evaluate its diagnostic value .Methods The expression of serum MALAT1 in 56 hospitalized TB patients , 35 latent TB infection (LTBI) individuals and 40 healthy controls were detected by real-time quantitative PCR .Serum levels of MALAT1 before and 3 ,6 months after anti-TB therapy in 16 TB patients were determined .Receiver operating characteristic (ROC) curve analysis was used to evaluate the sensitivity and specificity of serum MALAT 1 .The comparison between two groups was performed by Student t-test , and the comparison among three groups was performed with one-way analysis of variance test .Results Serum level of MALAT1 in TB patients was (2 .10 ± 1 .05) ,which was significantly higher than those in LTBI individuals (1 .16 ± 0 .51) and healthy controls (1 .02 ± 0 .44 ,F= 28 .53 ,P< 0 .01) .The MALAT1 level in TB patients with positive sputum smear was significantly higher than that in patients with negative sputum smear (2 .42 ± 1 .03 vs 1 .43 ± 0 .74 ,t= 2 .66 ,P< 0 .01) .Compared with pre-treatment (2 .28 ± 0 .79) ,the serum MALAT 1 levels decreased significantly in 3 months (1 .35 ± 0 .39) and 6 months (1 .05 ± 0 .30) after anti-TB therapy (t= 4 .33 ,6 .05 ;both P< 0 .01) .The area under the curve (AUC) of serum MALAT1 was 0 .821 , with sensitivity and specificity of 0 .732 and 0 .850 , respectively .Conclusion The expression of MALAT1 is up-regulated in TB patients , and could be used as potential novel biomarkers for the diagnosis of TB .
7.Expression of thrombospondin-1 in vaginal wall of bilateral ovariectomy rats
Fangyi ZHU ; Li HONG ; Mao CHEN ; Ya XIAO ; Xiaoyu HUANG ; Liying CHEN
China Modern Doctor 2023;61(34):1-4,8
Objective To explore the possible role of thrombospondin-1(THBS1)in the excessive fibrosis of vaginal wall induced by estrogen deficiency in rats,the morphological structure of collagen fibers and the expression of THBS1 in the vaginal wall were detected in the estrogen deficiency model of rats.Methods Twenty-four SD rats aged 3 months without delivery were randomly divided into sham operation group and experimental group,with 12 rats in each group.After 12 weeks of modeling,the rats were killed and the vaginal walls were taken for analysis.Masson staining was used to observe the morphological and structural changes of collagen fibers in vaginal wall of rats.Immunohistochemical staining and Western blotting were used to detect the expression level of THBS1 protein.Results After 12 weeks of modeling,the uterine atrophy of experimental group was obvious,the increase of body mass was significantly higher than that of sham operation group,and the level of estradiol was significantly lower than that of sham operation group(P<0.01).Compared with the sham operation group,the upper cortex of vaginal wall of experimental group was significantly atrophy,the smooth muscle bundles were thinner,the muscle gap was wider,the collagen fiber deposition in lamina propria and muscle layer was increased,and the arrangement and distribution were disordered and fragmented.THBS1 expression in vaginal wall of experimental group was significantly higher than that of sham operation group(P<0.05).Conclusion Estrogen deficiency may mediate excessive fibrosis of vaginal wall by upregulating THBS1 expression,thereby damaging the biomechanical properties of vaginal wall and leading to an increased susceptibility to pelvic organ prolapse development.
8.A Risk Prediction Model for Invasive Fungal Disease in Critically Ill Patients in the Intensive Care Unit.
Fangyi LI ; Minggen ZHOU ; Zijun ZOU ; Weichao LI ; Huang HUANG ; He ZHIJIE
Asian Nursing Research 2018;12(4):299-303
PURPOSE: Developing a risk prediction model for invasive fungal disease based on an analysis of the disease-related risk factors in critically ill patients in the intensive care unit (ICU) to diagnose the invasive fungal disease in the early stages and determine the time of initiating early antifungal treatment. METHODS: Data were collected retrospectively from 141 critically ill adult patients with at least 4 days of general ICU stay at Sun Yat-sen Memorial Hospital, Sun Yat-sen University during the period from February 2015 to February 2016. Logistic regression was used to develop the risk prediction model. Discriminative power was evaluated by the area under the receiver operating characteristics (ROC) curve (AUC). RESULTS: Sequential organ failure assessment (SOFA) score, antibiotic treatment period, and positive culture of Candida albicans other than normally sterile sites are the three predictors of invasive fungal disease in critically ill patients in the ICU. The model performs well with an ROC-AUC of .73. CONCLUSION: The risk prediction model performs well to discriminate between critically ill patients with or without invasive fungal disease. Physicians could use this prediction model for early diagnosis of invasive fungal disease and determination of the time to start early antifungal treatment of critically ill patients in the ICU.
Adult
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Candida albicans
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Critical Care*
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Critical Illness*
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Early Diagnosis
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Humans
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Intensive Care Units*
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Logistic Models
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Retrospective Studies
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Risk Factors
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ROC Curve
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Solar System
9.Clinical value of the plasma circular RNA hsa_circ_0009024 in the diagnosis and treatment of pulmonary tuberculosis
Zikun HUANG ; Qingshui HUANG ; Qing LUO ; Fangyi YAO ; Yiping PENG ; Jianqing XU ; Lu ZHANG ; Junming LI
Chinese Journal of Laboratory Medicine 2018;41(5):399-404
Objective Detecting plasma level of circular RNA(circRNA)hsa_circ_0009024 in pulmonary tuberculosis(TB)patients,and evaluating its diagnostic value for TB.Methods From January 2016 to December 2016, a hosptial-based, case-control study was performed, which include 90 untreated active pulmonary tuberculosis patients(TB group),75 healthy people(healthy control)and 84 patient with other diseases(other disease group).Plasma level of circRNA hsa_circ_0009024 was detected by real-time quantitative polymerase chain reaction.Furthermore, the 90 patients with TB were divided into different subgroups according to cavity formation and the lung fields involvement: patients without lung cavity(55 cases)vs those with lung cavity(35 cases),patients with involvement of <2 lung fields(49 cases)vs≥2 lung fields(41 cases).Plasma levels of hsa _circ_0009024 of 41 TB patientswere monitored andcomparedbefore and after 3 months anti-TB therapy.The sensitivity and specificity of plasma hsa_circ_0009024 were analyzed by using the receiver operating characteristic(ROC)curve.The comparison between two groups was performed with Mann-Whitney U test and the comparison among multigroupswas conducted with Kruskal-Wallis H test.Results Plasma levels of hsa_circ_0009024 in TB patients[1.98(1.42, 2.71)]were significantly higher than healthy controls[1.03(0.78,1.33)]and other disease groups[1. 13(0.77,1.51)](H=76.58,P<0.0001).Plasma levels of hsa_circ_0009024 in cavity pulmonary TB patients were higher than pulmonary TB patients without cavity(U=392.50,P<0.0001).Plasma levels of hsa_circ_0009024 in TB patients with involvement of ≥2 lung fields were higher than <2 lung fields(U=590.50,P=0.0008).As compared to pre -treatment[2.01(1.41, 2.71)], the plasma hsa_circ_0009024 levels decreased significantly in 3 months[1.22(0.85,1.47)](U=292.50,P<0.0001)after anti-TB therapy.The area under the receiver operating characteristics curve(AUC)of plasma hsa_circ_0009024 in discriminating the patients with TB from normal controls, pneumonia patients and lung cancer patients were 0.841 and 0.811, respectively.Conclusion The hsa_circ_0009024 can be used as a potential biomarker in TB diagnosis and monitoring.
10.Advice on the rationalized layout of outpatient clinics in a wound repair department
Ming ZHOU ; Chunlan WANG ; Jiajun TANG ; Yiwen NIU ; Yingkai LIU ; Yechen LU ; Lifang HUANG ; Jingqi ZHOU ; Fangyi WU ; Xian MA
Chinese Journal of Burns 2021;37(7):666-667
According to a document issued by the General Office of National Health Commission, "one person, one diagnosis, and one room" is required in the process of outpatient consultation. However, the patient will need to go to another room for dressing change after the doctor checks the wound if sticking to the conventional layout of current wound repair specialist outpatient clinic in hospitals and following the regulation of "separation of diagnosis and treatment". To allow a patient walking back and forth with the exposed wounds to different clinics or going to another clinic for dressing change with the original dressing reapplied to the wound is against the regulation of nosocomial infection control and the principle of sterility. To ensure that the layout of the outpatient clinic in the wound repair outpatient department not only conforms to the principle of "one person, one diagnosis, and one room", but also meets the characteristics of the diagnosis and treatment process of chronic wounds, this paper proposes the layout of "large space and small partition" in the wound repair clinic.