Objective To investigate the role of pathogenic Leptospira interrogans lipopolysaccha-ride (L-LPS) and outer membrane proteins (L-OMP) in the apoptosis of mouse macrophages (J774A.1) and their association with Fas/FasL pathway .Methods Phenol-water extraction and Triton X-114 phase separation were used to extract L-LPS and L-OMP from L.interrogans serogroup icterohaemorrhagiae serovar Lai strain Lai, respectively.Polymyxin B ( PMB) and protease K ( PK) were used to treat L-LPS and L-OMP, respectively.J774A.1 cells were stimulated by L.interrogans strain Lai with or without ultraviolet inactivation.In parallel, the cells were stimulated by extracted L-LPS and L-OMP with or without PMB and PK treatments .The apoptosis and necrosis of J 774 A.1 cells before and after treatment were detected by flow cytometry.The siRNAs were used to silence the expression of Fas or FasL gene in J 774A.1 cells and their inhibitory effects were further validated by using real-time fluorescent quantitative RT-PCR.Flow cytometry was used to detect the effects of L-LPS or L-OMP on the apoptosis of J774A.1 cells with Fas or FasL gene-knockdown .Results L.interrogans strain Lai with or without ultraviolet inactivation could cause similar early apoptosis rates (47.1%and 55.6%) and late apoptosis/necrosis rates (7.6%and 7.9%).The ear-ly apoptosis rates of 1×105 J774A.1 cells were 40.4%and 34.0%after the treatment with 100 ng of L-LPS and 100 μg of L-OMP for 4 h.The late apoptosis/necrosis rates of the cells were 7.5%and 6.9%upon the treatments with L-LPS and L-OMP, respectively.However, the apoptosis or necrosis of the cells was not ob-served when using L-LPS and L-OMP pre-treated by PMB and PK, respectively.Silenced expression of Fas or FasL gene reduced the L-LPS-induced J774A.1 cells apoptosis (P<0.05), while decreased early apopto-sis rate of J774A.1 cells mediated by L-OMP was only observed in Fas gene-knockdown cells (P<0.05). Conclusion Both L-LPS and L-OMP can cause the Fas/FasL-associated apoptosis of macrophages , which is beneficial for L.interrogans to establish the productive infection in hosts .

0.05).The expression of TNF-? mRNA was significantly upregulated in the lower respiratory tract,C-7-T-5 spinal ganglia and corresponding spinal dorsal horn of the experimental asthmatic guinea pigs compared with the normal saline control group and the simple sensitized group(P

BACKGROUND:Embryonic stem cells (ESCs) serve as a major cell source for smooth muscle cells,but the heterogeneity of cells derived from ESCs result in difficulty to obtain high purity smooth muscle cells.OBJECTIVE:To construct a double expression vector of puromycin resistance (pac) gene and enhanced green fluorescence protein (EGFP) gene driven by smooth muscle specific SM22α promoter (pSM22α-PAC-IRES2-EGFP),in addition,to detect its availability and specificity in ESCs.DESIGN,TIME AND SETTING:The observational experiment of gene level was performed at the Cardiovascular Institute,General Hospital of Shenyang Military Region from April 2007 to September 2008.MATERIALS:ESCs line R1 with number SCRC-1011TM was purchased from American ATCC Company.The pSM22α-EGFP vector was constructed by our laboratory.And the pIRES2-EGFP,pSM2C and pSuper.basic vectors were purchased from Invitrogen Company.METHODS:SM22α promoter was cloned from pSM22α-EGFP by polymerase chain reaction.CMV promoter of pIRES2-EGFP vector was replaced by SM22 promoter to establish pSM22α-IRES2-EGFP.Pac gene,excised from pSM2C by HindⅢ/Clal digestion,was sub-cloned into pSuper.basic to establish pSuper-PAC.After BgⅢ/Accl enzyme digestion of pSuper-PAC,pac gene fragment was obtained,which was further sub-cloned into pSM22α-IRES2-EGFP to produce pSM22α-PAC-IRES2-EGFP.ESCs were transfected with pSM22α-PAC-IRES2-EGFP using lipofectamine.Positive clones were selected by G418 and induced to differentiate and further identified by amplification of pac gene by RT-PCR.Differentiated cells were immunostained by SM α-actin,and expression of SM α-actin and EGFP was observed simultaneously under fluorescence microscope.MAIN OUTCOME MEASURES:Sequencing result of pSM22α-PAC-IRES2-EGFP;Amplification of pac gene;EGFP expression;as well as SM α-actin immunostaining.RESULTS:Three segments of 261 bp,664 bp,and 5000 bp were obtained by HindⅢ/Clal digestion,which was coincident with expectation,and the sequencing results showed that pSM22α-PAC-IRES2-EGFP vector was successfully constructed.Amplification of pac gene identified 4 ESCs clones successfully transfected.After induction of differentiation,partial portion of differentiated cells expressed EGFP,accompanied by positively stained by SM α-actin antibody.CONCLUSION:pSM22α-PAC-IRES2-EGFP vector was successfully constructed.ESCs clones transfected with this vector expressed pac gene and EGFP gene,and the expression of EGFP is smooth muscle specific.

Objective:To observe the recent effects and toxicity of thermochemotherapy on malignant hydrothorax or hydroperitoneum,to evaluate the changes of the immunological functions,and to investigate the mechanism of thermochemotherapy.Methods:Fifty-two patients were treated with weekly intracavitary chemotherapy,and then combined with local endogenetic thermotherapy twice a week.As the control,another 50 patients received weekly intracavitary chemotherapy.The treatment lasted for two weeks and was followed by one-week rest,and then the recent effects and toxicity were observed.The T cell subset,NK cells and VEGF levels in serum,hydrothorax or hydroperitoneum were tested.Results:Overall response rates of the malignant hydrothorax were 86.9% vs 60.0%(P

Objective:Position emission tomography/computed tomography(PET/CT) is a new bio-imaging system which is combined metabolic with anatomic imaging.This study was to compare the accuracy of conventional staging methods(including computed tomography,ultrasound,magnetic resonance imaging,and detection of bone marrow) with that of PET/CT for lymphoma staging and re-staging. Methods:A total of 42 patients with lymphoma diagnosed by operation or biopsy,received conventional and PET/CT staging.The accuracy of these two methods and their impact on lymphoma staging were compared.Results:The accuracy of PET/CT scan was 95.2%(40/42),and that of conventional staging was 78.6%(33/42).The detection rates of internal lymph node were 97.1%(66/68) and 88.2%(60/68),respectively.The detection rates of outer lymph node were 91.7%(22/24) and 58.3%(14/24),respectively.Compared with conventional staging methods,7 cases were up-staged and 2 cases were down-staged by PET/CT,which led to the change of therapy in 8 cases.Conclusion:PET/CT scan is more sensitive and accurate than conventional staging methods in staging and restaging of lymphoma.

OBJECTIVETo compare the diagnostic value of antibodies to mutated citrullinated vimentin (MCV) and some associated autoantibodies in juvenile idiopathic arthritis and to further analyze the relation between antibodies and inflammatory markers.

METHODAntibodies to cyclic citrullinated peptides (CCP) and anti-MCV antibodies were detected by enzyme-linked immunosorbent assay (ELISA), antiperinuclear factor (APF) and antikeratin antibody (AKA) by indirect immunofluorescent assay, as well as rheumatoid factor (RF) by latex agglutination test in serum samples from 113 patients with JIA and 56 children without rheumatoid arthritis.

RESULT(1) The positive rate of anti-MCV antibodies, anti-CCP antibodies, and RF was 16.8%, 14.2%, and 21.2% in the JIA. In the other group, the positive rate was 2.2%, 2.2%, and 6.5%. There was a significant difference between the two groups (χ(2)=8.105, 6.337, 7.036, P<0.05). The positive rate of AKA and APF were not significantly different. The area under the ROC curve of anti-MCV antibodies, anti-CCP antibodies, RF, AKA, APF was 0.579, 0.561, 0.578, 0.539, 0.505. (2) The positive rate of anti-MCV antibodies and anti-CCP antibodies were higher than other antibodies. In the RF-positive polyarticular disease patients, they were higher than those in the other subtypes (P<0.05). Antibody levels were not significantly different (P>0.05) from other subtypes. (3) The swollen joint counts and tender joint counts had a low correlation to anti-MCV antibodies, anti-CCP antibodies, RF, AKA and APF. No correlation was found between ESR, CRP and anti-MCV antibodies, anti-CCP antibodies, RF, AKA and APF.

CONCLUSIONThe diagnostic value of anti-MCV antibodies is low for JIA. The positive rate of anti-MCV antibodies was higher than the other antibodies in the classification of JIA. There was a low correlation between anti-MCV antibodies, anti-CCP antibodies, RF, AKA, APF and swollen joint counts, tender joint counts.

Antibodies, Antinuclear ; blood ; Arthritis, Juvenile ; blood ; Arthritis, Rheumatoid ; Autoantibodies ; blood ; Biomarkers ; blood ; Child ; Enzyme-Linked Immunosorbent Assay ; Fluorescent Antibody Technique, Indirect ; Humans ; Peptides, Cyclic ; immunology ; ROC Curve ; Rheumatoid Factor ; blood ; Vimentin ; immunology

Objective: To investigate the refractive status of children and adolescents aged 4-18 years in Chengdu, and to analyze the epidemiological characteristics of refractive parameters. Methods: A cross sectional survey was conducted among 82 024 children and adolescents aged 4-18 years in Gaoxin District of Chengdu from August to September 2021. The prevalence of screening myopia, low vision rate, high myopia rate, refractive status and axial development were analyzed. Results: The prevalence of screening myopia in students aged 4-18 years was 40.42%(33 158/82 024). Low myopia was 24.51%(20 108/82 024), moderate myopia was 13.05%(10 703/82 024) and high myopia was 2.86%(2 347/82 024). The prevalence of screening myopia was 1.81% (233/12 848) in kindergarten, 34.44%(17 095/49 644) in primary school, 79.73%(9 738/12 214) in junior high school and 83.25% (6 092/7 318) in senior high school. The rate of visual impairment increased by year from the age of 4, and the rate of myopia increased most rapidly from 6 to 15 years old, the prevalenct of high myopia was compared between adjacent age groups: there were statistical differences between 9-15 years old were more likely to be nearsighted than boys( P <0.035 7). Significant differences in screening myopia between 5- and 7-17 year old groups,and in boys and girls.The median total diopter was 0.40 D at the age of 4 and developed to -2.90 D at the age of 18 . The diopter of girls aged 8-15 years was higher than that of boys, and there was statistical significance( Z=-2.53, -4.09, -5.67, -8.64, -5.56, -4.97, -2.52, -2.14, P <0.05). The axial length gradually increased with age, with the mean value of (22.31±0.59) mm at 4 years old and (24.91±1.00) mm at 18 years old. The mean corneal curvature did not change with age (43.19±1.47)D. Conclusion 6-15 years old is a especially critical period for myopia development. More efforts need to be taken to decrease the prevalence of myopia before 6 years old. Prevention of the development of high myopia should start before the age of 10. The prevalence of myopia in girls is higher than that in boys, more prevention and control of myopia should focus on girls.

Objective:To explore the clinical features and risk factors of systemic lupus erythematosus(SLE) concomitant with interstitial lung disease(ILD) in children.Methods:A retrospective analysis was performed.A total of 111 hospitalized children diagnosed with SLE in the Department of Rheumatology and Immunology, Children′s Hospital of Soochow University from February 2016 to November 2018 were selected as the research subjects and divided into the SLE-ILD group(18 cases) and the SLE-non-ILD group(93 cases)according to the lung high-resolution CT manifestations. T-test and Wilcoxon rank sum test were used to compare and analyze the general situation, clinical manifestations and laboratory results.Multivariate Logistic regression was used to analyze the risk factors of SLE-ILD. Results:The prevalence of SLE-ILD was 16.2%(18/111 cases). There were significant differences between the SLE-ILD group and the SLE-non-ILD group in the course of disease [14.00 (12.00-24.25) months vs.1.00(1.00-2.00) months], the incidence of serositis [55.6%(10/18 cases) vs.8.6%(8/93 cases)], post-activity shortness of breath [83.3%(15/18 cases) vs.25.8%(24/93 cases)], nervous system damage [27.8%(5/18 cases) vs.6.5%(6/93 cases)], cardiovascular system damage [38.9%(7/18 cases) vs.9.7%(9/93 cases)], the occu-rrence of increased erythrocyte sedimentation rate [66.7%(12/18 cases) vs.31.2%(29/93 cases)], the decreased C 3[88.9%(16/18 cases) vs.62.4%(58/93 cases)], positive anti neutrophil cytoplasmic antibody (ANCA) [88.9%(16/18 cases) vs.18.3%(17/93 cases)], positive anti-Sm antibody [61.1%(11/18 cases) vs.15.1%(14/93 cases)] and anti ribonucleoprotein antibody (anti RNP antibody)[66.7%(12/18 cases) vs.16.1%(15/93 cases)](all P<0.05). Logistic regression analysis demonstrated that serositis( OR=30.535, 95% CI: 2.167-430.336, P=0.011), shortness of breath after exercise( OR=55.115, 95% CI: 1.117-2 579.852, P=0.041), positive ANCA( OR=65.090, 95% CI: 4.488-944.071, P=0.002) and positive anti-RNP antibody( OR=10.007, 95% CI: 1.362-73.500, P=0.024) were risk factors for SLE-ILD. Conclusions:The longer the course of SLE, the higher the incidence of ILD; serositis, shortness of breath after exercise, positive ANCA and positive anti RNP antibody may be risk factors for SLE-ILD.

As a designated hospital, Tongji Hospital shoulders the task of diagnosis and treatment of numerous patients of the disease. Based on the medical cloud platform, the hospital has initiated a regional remote diagnosis center; leveraging its IT system, the hospital initiates its epidemic prevention and management mechanism, sets up a self-service system for patients at the fever clinic, launches its online diagnosis and treatment services, and establishes a hospital epidemic supervision platform. By strengthening the informational support needed for epidemic prevention and control, the hospital has enhanced its efficiency of epidemic prevention and control, reducing the risk of cross-infection, and ensuring data security. Its experiences offer references for informationization support for other regions and hospitals in China.

December of 2019 witnessed the outbreak of new coronavirus pneumonia in Wuhan city and a few localities. As a designated hospital, Tongji Hospital is designated as a hospital for the diagnosis and treatment of numerous patients of such a disease. Based on the medical cloud platform, the hospital has initiated a regional remote diagnosis center; based on its IT system, the hospital to operate its epidemic prevention and management mechanism, set up the self-service system for patients at the fever clinic, launched its online diagnosis and treatment services, and established a hospital epidemic supervision platform. By strengthening the informational support needed for epidemic prevention and control, the hospital has enhanced its efficiency of epidemic prevention and control, reducing the risk of cross-infection, and ensuring data security. Its experiences offer references for informationization support for other regions and hospitals in China.