1.Effects of RNA Interference Combined with Ultrasonic Irradiation and SonoVue Microbubbles on Expression of STAT3 Gene in Keratinocytes of Psoriatic Lesions
RAN LI-WEI ; WANG HAO ; LAN DONG ; JIA HONG-XIA ; YU SI-SI
Journal of Huazhong University of Science and Technology (Medical Sciences) 2017;37(2):279-285
The most effective sequence of small interfering RNA (siRNA) silencing STAT3 of psoriatic keratinocytes (KCs) was screened out,and the effects of the most effective siRNA combined with ultrasonic irradiation and SonoVue microbubbles on the expression of STAT3 of KCs and the dose-and time-response were investigated.Three chemically-synthetic siRNAs targeting STAT3 carried by Lipofectamine 3000 were transfected into KCs,and the effects on STAT3 expression were detected,then the most effective siRNA was selected for the subsequent experiments.The negative controls of siRNA (siRNA-NC) labeled with Cy3 carried by Lipofectamine 3000 combined with ultrasonic irradiation and SonoVue microbubbles were transfected into KCs,then the optimal parameters of ultrasonic irradiation were determined.The most effective siRNA carried by Lipofectamine 3000 combined with ultrasonic irradiation at the optimal parameters and SonoVue microbubbles was transfected into KCs,and the dose-and time-response of RNA interference was determined.The effect of RNA interference by the most effective siRNA at the optimal time and dose carried by Lipofectamine 3000 combined with ultrasonic irradiation and SonoVue microbubbles (LUS group) was compared with that only carried by Lipofectamine 3000 (L group).The results showed that siRNA-3 achieved the highest silencing efficacy.0.5 W/cm2 and 30 s were selected as the parameters of ultrasonic irradiation.The siRNA-3 carried by Lipofectamine 3000 combined with ultrasonic irradiation and SonoVue mierobubbles could effectively knock down the STAT3 expression at mRNA and protein levels in dose-and time-dependent manners determined at 100 nmol/L with maximum downregulation on mRNA at 48 h,and on protein at 72 h after transfection.The LUS group achieved the highest silencing efficacy.It was concluded that siRNA-3 carried by Lipofectamine 3000 combined with ultrasonic irradiation and SonoVue microbubbles could effectively knock down the STAT3 expression in psoriatic KCs,and the optimized transfection condition and the sequence of siRNA-3 could serve for further research on gene therapy of psoriasis.
2.The Reliability Analysis of the Chinese Version of Speech Intelligibility Rate (SIR)
Yu WANG ; Tao PAN ; Si MI ; Furong MA
Journal of Audiology and Speech Pathology 2013;(5):465-468
Objective To discuss the application of Chinese version of Speech intelligibility rating (SIR) in e-valuating the speech intelligibility of Chinese children with hearing disorder ,and analyze its scorer reliability .Meth-ods 50 children with cochlear implants or hearing aids were interviewed using SRI Chinese version .Each child was rated by a parent and a speech and language therapist .The reliability of scores was analyzed with Spearman correla-tion coefficient ,Kendall's coefficient of concordance and Kappa coefficient .Results The complete agreement was foud in 34 cases(68% ) ,with a disagreement of one category in 14 cases(28% ) ,and a disagreement of 2 categories was found in 2 cases (4% ) .Spearman correlation coefficient was 0 .885 ,Kendall's coefficient of concordance was 0 .801 ,Kappa coeffi-cient was 0 .593 ,the result was statistically significant (P<0 .001) .Conclusion SRI Chinese version has good reliability and is helpful in evaluating Chinese speech producing ability of children with hearing disorder in everyday life .
3.Clinical analysis of eight cases of adult-onset Still's disease
He'nan SI ; Shanshan LI ; Hong WANG ; Hua ZHENG ; Yan YU ;
Chinese Journal of Dermatology 2017;50(9):654-657
Objective To investigate characteristic skin lesions and typical histopathological changes of adult-onset Still's disease(AOSD) for its early diagnosis and treatment.Methods Clinical data were collected from 8 patients with AOSD,and analyzed retrospectively.Results All the patients had transient rashes and persistent papules/plaques during the course of disease.Of the 8 patients,1 had urticaria-like rashes,3 had dermatomyositis-like rashes,and 1 had prurigo pigmentosa-like rashes.Biopsies were carried out at the sites where transient rashes or persistent papules/plaques occurred.Histopathological findings showed necrotic keratinocytes in the upper prickle cell layer,and perivascular infiltration of neutrophils and lymphocytes in the upper dermis.Conclusion The skin lesions and histopathological changes of AOSD are characteristic,which can provide clues to the early diagnosis of AOSD.
4.Transplantation of bone marrow-derived mesenchymal stem cells improves acute lung injury in mice induced by LPS
Lei SUN ; Xiaoming WANG ; Qin SI ; Xiaohong YU ; Yu LIN ; Jin QIU ; Hengyi GUO ; Qixia WU
Basic & Clinical Medicine 2009;29(12):1233-1238
Objective To study the potential efficacy of transplanted bone marrow-derived mesenchymal stem cells (MSCs) in treating and repairing the acute lung injury in animal models. Methods MSCs were isolated from mouse bone marrow, cultrued and amplified in vitro. The lipopolysaccharide (LPS) was inhaled through postnasal tract to cause acute lung injury in mice and the MSCs labeled by Brdu were administrated via vein into the mice. The migration and differention of the cells were identified by immunostaining and double immunostaining. The pathological changes, pulmonary edema index and the content of IL-1β in lung homogenate were used to accese the therapeutical effect of MSCs. Results The cultured MSCs dispalyed a positive CD44 and a negative CD34. The Brdu-labeled cells were detected in the lungs of the recipient 4 days after transplantation, indicating its origin of MSCs. Theses cells also exhibited characteristics of aveolar epithelials, expressing the cytokeratin-the marker of epithelium. Compared with the injuried ones, the mice treated with MSCs showed a decreased pulmonary edema in-dex and IL-1β content in the lung homogenate. Conclusion These data suggest a therapeutical effects of MSCs in treating and repairing the mouse acute lung injury.
5.Proliferation of retinal pigment epithelial cells induced by (R,R)-XY-10 and (S,S)-XY-10 and their action mechanisms
Yu-Wen, CHENG ; Yu-Liang, WANG ; Yi-Hua, ZHANG ; Si-Xun, PENG ; George C Y CHIOU
International Eye Science 2009;9(9):1641-1645
AIM: To investigate the mechanism of proliferation effect induced by (R,R)-XY-10 and (S,S)-XY-10 on retinal pigmented epithelial cells(ARPE-19).METHODS: Human retinal pigmented epithelial cells(ARPE-19) and human umbilical vein endothelial cells (HUVECs) were used to investigate the effect of (R,R)-XY-10 and (S,S)-XY-10 on cell growth,and their mechanisms of proliferative action by using ERK、 AKT、PI3K、Protein kinase C (PKC)and Nitric oxide synthase (NOS) inhibitors.RESULTS: (R,R)-XY-10 and (S,S)-XY-10 dose-dependently increased ARPE-19 cell proliferation,but not on HUVECs. When treated with proliferative inhibitors,H7(5μmol/L)、hypericin(20μmol/L)、PD98059(2μmol/L)、LY294002(50μmol/L)、SH-5 (10μmol/L) and L-NAME (100μmol/L),the proliferative effect was reduced by H7、hypericin、PD98059 and LY294002,but not by SH-5 and L-NAME.CONCLUSION: (R,R)-XY-10 and (S,S)-XY-10 can induce cell proliferation through MAPK and PI3K dependent pathway. KEYWORDS: age-related macular degeneration; (R,R)-XY-10; (S,S)-XY-10; ARPE-19 cells; human umbilical vein endothelial cells; proliferation
6.Antagonistic Mechanisms and Related Properties of Strain H5 Against Black-rot Disease of Dracaena sanderiana
Man CHEN ; Chi LI ; Yi-Si QIU ; Jian-Yu WANG ; Li YU ;
Microbiology 1992;0(04):-
A strain no.H5 isolated from Rhizophora stylosa Griff in Zhanjiang had a good antagonistic activity against Aspergillus niger,pathogen of Dracaena Sanderiana black-rot disease.It was identified as Bacillus licheniformis.Dual culture,mycelium growth rate and inhibitory zones were used to test the effect. Strong inhibition was shown against A.niger.Inhibitory ratios of H5 germ-free fermented filtrate on mycelium growth and conidial germination were 91.9%and 100%respectively.In addition,mycelia on the edge of antagonistic band became abnormal and over-branching.Meanwhile,a lot of vesicles appeared on the surface.When treated with heat,acid and alkali,the filtration of H5 was always with stable activity.Precipitate in 55%saturated ammonium sulfate dissolved in phosphate buffer solution maintained most of the activity after high pressure steam sterilization for 25 minutes.It was preliminarily considered as a kind of heat resistant protein.
7.Immunogenicity of a recombinant chimeric antigen using Aβ1-15 epitope fused to a T helper epitope
Si LIU ; Meng ZHAO ; Wenhui XU ; Yunzhou YU ; Shuang WANG ; Weiyuan YU ; Qing XU ; Zhiwei SUN
Military Medical Sciences 2014;(1):44-47,52
Objective To explore the immunogenicity of recombinant chimeric 6Aβ15-T including the Aβ1-15 epitope and a T-helper epitope formulated with different adjuvants and to evaluate its feasibility as a candidate vaccine for Alzheimer disease (AD).Methods The recombinant chimeric antigen 6Aβ15-T formulated with Al adjuvant, Freund′s adjuvant or MF59 adjuvant was administered to two strains of mice .The 6Aβ15-T-immunized group without adjuvants ( Mock) and non-immunized group (Control) were included in this study as control groups .The specific antibody and cellular immune response of the chimeric antigen were evaluated .Results In BALB/c strain mice, three types of adjuvants could substan-tially boost the immunogenicity of chimeric antigen 6Aβ15-T and produce a high level of specific-Aβ(β-amyloid) antibod-ies.In C57BL/6 strain mice, the existence of adjuvants enhanced the immune response of 6Aβ15-T antigen, but the mice in Mock group also produced a strong antibody response .In two strains of mice, prevalence of anti-AβIgG1, which was an indicator of Th2 polarization, was observed in the 6Aβ15-T-immunized mice.Additionally, the Al adjuvant induced a high-er level of IgG1 antibody titers, and the ratio of IgG1/IgG2a was the largest.As expected, the 6Aβ15-T antigen formulated with or without adjuvants induced PADRE-specific, but not Aβ42-specific T cellular immune response .Conclusion The 6Aβ15-T antigens formulated with different types of adjuvants could induce strong Th 2-polarized Aβ42-specific antibody re-sponses without activating self-reactive Aβ42-specific T cells in two strains of mice .The results suggested that the recombi-nant chimeric antigen 6Aβ15-T is a good candidate vaccine for AD .
8.Analysis of surveillance results of drinking-water-borne endemic fluorosis in Jiangsu Province in 2009
Chang-liang, SHU ; Cai-sheng, WANG ; Yang, WANG ; Yu-ting, XIA ; Si-hong, CHEN
Chinese Journal of Endemiology 2013;32(6):662-667
Objective To investigate the development trend of drinking-water-borne endemic fluorosis in Jiangsu Province,and to provide the basis for further prevention and treatment of the disease.Methods In 2009,eight major counties were chosen,and in each county all diseased villages were classified into light,moderate and severe disease types according to water fluorine content based on historical data,and one village was chosen from each type.In monitoring villages with improved water,one source water and three tap water samples were collected,respectively.Five water samples were collected in water unimproved monitoring villages according to water well locations of the east,the west,the south,the north and the center.The fluorine content in water was determined according to the Standard Testing Methods for Drinking Water (GB/T 5750-2006).Children aged 8 to 12 were examined for dental fluorosis by Dean method.Residents over the age of 16 were examined for clinical osteofluorosis,and two monitoring counties were chosen,then one village was respectively chosen in each county,and clinically diagnosed patients with skeletal fluorosis were examined again by X-ray.Both clinical and X-ray diagnosis were on the basis of Diagnostic Criteria of Endemic Skeletal Fluorosis (WS 192-2008).Urine samples of 30 children aged 8 to 12 and of 20 adults over the age of 16 were randomly collected and urinary fluoride was determined by F-ion selective electrode method(WS/T 89-1996).Results Of all the 24 villages of 8 counties,20 villages were water improved,and water-improvedprojects ran normally in 18 villages,while scrapped in the rest 2 villages.One hundred and two samples were tested,and the mean of water fluoride in water-improved villages was 1.10 mg/L,while in water unimproved villages and villages with water improved projects scrapped was 1.90 mg/L.The prevalence of dental fluorosis was 42.51%(854/2009).The prevalence of clinical skeletal fluorosis was 23.23% (2024/8713) and the X-ray detection rate was 32.00% (24/75).Six hundred and sixty-four urine samples of children were determined,and geometric mean of urinary fluorine was 1.59 mg/L,while 370 adult urine samples were determined,and geometric mean of urinary fluorine was 2.20 mg/L.Conclusions Endemic fluorosis in Jiangsu Province has not been fully controlled and there are signs of recovery.We must pay attention to water improvement measures to reduce fluoride and the management and maintenance of water improvement projects,and further strengthen the prevention and control of endemic fluorosis.
9.Spontaneous Cervical Epidural Hematomas in Mild Cervical Spondylotic Myelopathy Patients: An Analysis of 8 Cases
TANG SI-CHENG ; WANG YAN ; WANG YU ; YANG LEI ; CHEN JUN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2017;37(2):248-252
Spontaneous cervical epidural hematoms (SCEH) complicated with mild cervical spondylotic myelopathy (CSM) is a rare but emerging condition.Early diagnosis and treatment are important for good outcomes.This study aimed to investigate the clinical characteristics of this condition and to discuss the optimal treatment.The clinical data from 8 patients with SCEH plus CSM who were divided into two groups by treatment methods were retrospectively analyzed.The neurological function of the patients was assessed by Japanese Orthopedic Association (JOA) score before and after the surgical operations.Other factors were reviewed with medical records.Among them,4 out of the 8 patients underwent emergency surgery,and the rest 3 patients experienced an initial conservative treatment and ultimately received a laminectomy.We found that the Frankel Scale scores in most of the surgical patients were increased after surgery (6/7,85.7%).However,the JOA scores at the 6th month after onset were even lower than those before onset in 3 of the operative cases,and those in the patients who were given conservative treatment showed no significant change.It was concluded that some patients with SCEH and CSM treated with a timely operation may obtain relief from their previous CSM symptoms.However,the final neurological deficits of these patients were closely related to the progressive interval which refers to the hours between the initial onset and the occurrence of new neurological deficits or mild CSM deterioration,no matter whether they accept the operation.We found the crucial progressive interval may be in 9 h.Early MRIand prompt neurosurgical intervention are also important to improve the neurological deficits.
10.The factor analysis of the incidence of pneumothorax after CT-guided transthoracic needle aspiration biopsy
Tao ZHONG ; Hong-Guang YU ; Yong WANG ; Si-Fu YANG ; Xiao-Xuan WANG ;
Chinese Journal of Radiology 2000;0(11):-
Objective To analyze the impact of multiple factors on the incidence of pneumothorax associated with CT-guided transthoracic needle aspiration biopsy.Methods The sign of pneumothorax after 162 cases(lesion diameter from 1cm to 6cm)CT-guided transthoracic needle aspiration biopsy was observed and its relationship with multivariate factors were analyzed by multivariate logistic regression model.Results Thirty-two cases presented pneumothorax accounting for 19.8%.Single variate analysis showed that the sign of pneumothorax ralated to intercurrent COPD,distance from lesion and chest wall,needle dwelling time and lesion diameter.67 patients of intercurrent COPD with postoperative pneumothorax occurred in 22 cases (32.8%);With respect to those having lesions close to the chest wall(48 cases),and the cases with the distance between the chest wall and lesions less than 2cm(55 cases)and greater than 2cm(59 cases), the postoperative pneumothorax occurred in 0,14(25.5%),18(30.5%)cases respectively;For those patients with needle in the chest residence time of less than 10 minutes(82 cases),10—20 minutes (51 cases),more than 20 minutes(28 cases)after the occurrence of pneumothorax were 8(9.6%), 10(19.6%),14(50%)cases respectively;In contrast,those with lesion diameter less than 2 cm (65 cases),2—4cm(52 cases),more than 4cm(45 cases)were 19(29.2%),8(15.4%) and 5(11.1%)respectively.The multivariate logistic regression analysis showed that the prior three factors were risk factors of pneumothorax(OR=4.652,4.030,2.855 respectively).Conclusions To avoid the pneumothorax,caution must be taken with respect to CT-guided transthoracic needle aspiration biopsy, patients with intercurrent COPD,long distance between lesion and chest wall,and smaller lesion diameter. For operation the needle dwell time within thorax should be minimized.