1.Biological properties of concentrated growth factor combined with bone marrow mesenchymal stem cell sheet and its effect on bone defect repairment
Jianhong SHI ; Yuanye TIAN ; Kai CHEN ; Gao SUN ; Guomin WU
Journal of Jilin University(Medicine Edition) 2024;50(6):1535-1546
Objective:To discuss the effect of concentrated growth factor(CGF)on the performance of bone marrow mesenchymal stem cells(BMSCs)sheets,and to clarify the role of CGF-containing composite cell sheets(CS)in the bone defect repairment.Methods:In in vitro experiments,the BMSCs were isolated and cultured from two 3-week-old SD rats;Alizarin Red S and Oil Red O staining were used to identify the osteogenic and adipogenic capabilities of BMSCs;CGF liquid extracts(CGFe)was prepared from three 3-week-old SD rats.The cells were divided into control group,traditional CS(BMSC-CS)group,and CGF-containing composite CS(CGF/BMSC-CS)group.The morphology of the CS in two groups was observed by HE staining.Alizarin Red and alkaline phosphatase(ALP)staining were used to detect the osteogenic differentiation of the CS in various groups;cell scratch assay was used to detect the migration abilities of the cells in various groups;real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the mRNA expression levels of ALP,collagen are type 1(COL-1),Runt-related transcription factor 2(RUNX2),and osteocalcin(OCN)in the cells in various groups.In in vivo experiments,15 SD rats were randomly divided into control group,BMSC-CS group,and CGF/BMSC-CS group;micro computed tomography(Micro-CT)was used to detect the bone formation parameters in skull defects of the rats in various groups;HE staining and Masson staining were used to observe the morphology of skull defect tissue of the rats in various groups.Results:The third-generation BMSCs were spindle-shaped,closely arranged,and grew in a vortex cluster.The Alizarin red staining results showed obvious calcium nodules,and the Oil red O staining showed red lipid droplets,confirming the cells'ability to undergo osteogenic and adipogenic differentiation.The CS were white and semi-transparent,with slightly curled edges.The peeled CS were irregularly curled and wrinkled.Compared with BMSC-CS group,the CS in CGF/BMSC-CS group were whiter,less transparent,significantly increased in thickness and extensibility,less prone to breakage,and had a certain degree of stickiness and plasticity.The HE staining results showed that compared with BMSC-CS group,the number of the cells of CS in CGF/BMSC-CS group was increased,with denser arrangement and more abundant extracellular matrix(ECM),which wrapped and connected the cells to form an integral sheet-like structure.The Alizarin red and ALP staining results showed that compared with control group,the ALP activity and mineralization uplift value of CS in BMSC-CS group were significantly increased(P<0.05);compared with control group and BMSC-CS group,the number of osteoblasts and red mineralized nodules in the CS in CGF/BMSC-CS group was significantly increased,with obvious deepening of the staining,increased positive area,and the ALP activity and mineralization uplift value were significantly increased(P<0.05).Compared with BMSC-CS group,the ALP activity and mineralization uplif value of the CS in CGF/DMSC-CS group were increased(P<0.05).The cell scratch assay results showed that after 24 of culture,compared with control group,the migration rates of the cells in BMSC-CS group and CGF/BMSC-CS group were significantly increased(P<0.05).Compared with BMSC-CS group,the migration rate of the cells in CGF/BMSC-CS group was significantly increased(P<0.01).After 48 h of culture,compared with control group,the migration rate of the cells in CGF/BMSC-CS group was significantly increased(P<0.05).The RT-qPCR results showed that compared with control group,the expression levels of COL-1 and OCN mRNA in the cells in BMSC-CS group were significantly increased(P<0.01),and the expression levels of ALP,COL-1,OCN,and RUNX2 mRNA in the cells in CGF/BMSC-CS group were significantly increased(P<0.01).Compared with BMSC-CS group,the expression levels of ALP,COL-1,and OCN mRNA in the cells in CGF/BMSC-CS group were significantly increased(P<0.01).The Micro-CT detection results showed that in control group,the boundary of the rat skull defect area was clear,with almost no new bone formation.In BMSC-CS group,a small amount of new bone formed only at the edge of the bone defect in skull of the rats,with a significant gap in the central area of the defect.In CGF/BMSC-CS group,new bone formed along the edge of the bone defect towards the central area in skull of the rats,repairing most of the bone defect.Compared with control group,the bone volume(BV)and trabecular number(Tb.N)of the rats in BMSC-CS group were significantly increased(P<0.05);the bone volume(BV),bone volume fraction[BV/tissue volume(TV)],trabecular thickness(Tb.Th),and trabecular number(Tb.N)in skull of the rats in CGF/BMSC-CS group,were significantly increased(P<0.05).Compared with BMSC-CS group,the BV,BV/TV,Tb.Th,and Tb.N in skull of the rats in CGF/BMSC-CS group were significantly increased(P<0.01).The HE and Masson staining observation showed that in control group,almost no new bone formed in the skull defect tissue of the rats,with only a large amount of collagen fibers connecting the two sides of the bone ends.In BMSC-CS group,a small amount of new bone formed only at the edge of the bone defect in skull tissue of the rats,with the central area of the defect containing dense collagen fibers connected to the newly formed bone at the defect edge.In CGF/BMSC-CS group,new bone tissue could be seen at the edge of the bone defect,and bone islands formed in the central area of the defect,surrounded by osteocytes and a large amount of collagen fibers.The Masson staining observation results showed that the cytoplasm and osteoid were red,and the collagen was blue.In CGF/BMSC-CS group,newly formed osteoid was observed in skull defect tissue of the rats,with the highest amount of new bone formation.Conclusion:CGF can promote the osteogenic differentiation and increase the richness of ECM in BMSCs sheets.CGF-containing composite CS can efficiently repair skull defects of the rats and serve as an ideal and safe material for promoting the bone regeneration.
2.Therapeutic effect of resveratrol on osteoarthritis of temporomandibular joint and its mechanism
Gao SUN ; Jing HE ; Qi ZHAO ; Jianhong SHI ; Zhiling LIAO ; Yuanye TIAN ; Guomin WU
Journal of Jilin University(Medicine Edition) 2024;50(6):1547-1556
Objective:To discuss the therapeutic effect of resveratrol on the temporomandibular joint osteoarthritis(TMJOA),and to clarify the related mechanism.Methods:Forty-five SD rats were randomly divided into control group,model group,and resveratrol group,and there were 15 rats in each group.The rats in model group and resveratrol group were intra-articularly injected with 50 μL of 20 g·L-1 monosodium iodoacetate(MIA)to set TMJOA rat models,while the rats in control group were injected with an equal volume of normal saline.Three weeks after modeling,the rats in resveratrol group received an injection of 80 μL resveratrol solution,once a week for three weeks,while the rats in control and model groups were injected with an equal volume of normal saline.Micro-computed tomography(Micro-CT)system was used to detect the condyle structure and the bone volume fraction(BV/TV),trabecular thickness(Tb.Th),trabecular spacing(Tb.Sp),and trabecular number(Tb.N)of the rats in various groups were calculated;HE staining and toluidine blue staining were used to observe the pathomorphology of temporomandibular joint(TMJ)tissue of the rats in various groups;immunohistochemistry was used to detect the expression levels of SRY-related HMG box(SOX)-9,matrix metalloproteinase(MMP)-13,silent information regulator(Sirt)1,phosphatidylinositol 3-kinase(PI3K),phosphorylated protein kinase B(p-Akt),and phosphorylated mammalian target of rapamycin(p-mTOR)in TMJ tissue of the rats in various groups;real-time quantitative PCR(RT-qPCR)method was used to detect the expression levels of SOX-9,MMP-13,Sirt1,PI3K,mTOR,and Akt mRNA in TMJ tissue of the rats in various groups.Results:Three weeks after modeling,condylar bone was destructed,the surface was roughness,and continuity interruption were observed,indicating TMJOA model of the rats was established successfully.The Micro-CT system results showed that the condylar surface of the rats in control group was smooth and regularly shaped,with continuous bone texture;the rats in model group had significant condylar destruction,disrupted continuity,surface roughness,and varying degrees of bone defects;the rats in resveratrol group showed alleviated condylar lesions and improved appearance.Compared with control group,the BV/TV and Tb.Th of the rats in model group were significantly decreased(P<0.05),and Tb.Sp was significantly increased(P<0.05);compared with model group,the BV/TV and Tb.Th of the rats in resveratrol group were significantly increased(P<0.05),and the Tb.Sp was significantly decreased(P<0.05).The HE staining results showed clear layers and orderly chondrocyte arrangement in condyle of the rats in control group;the rats in model group showed rough uneven surface,obvious defects,and typical TMJOA features;the rats in resveratrol group showed slightly rough surface with generally clear layers and orderly arranged cells.The toluidine blue staining results showed distinct blue-purple staining of chondrocytes in hypertrophic layer of the rats in control group;pale staining or even loss of staining in some areas of the rats in model group;and distinct and relatively uniform staining in hypertrophic layer of the rats in resveratrol group.The immunohistochemistry results showed that compared with control group,the expression levels of MMP-13,PI3K,p-Akt,and p-mTOR proteins in TMJ tissue of the rats in model group were significantly increased(P<0.05),while the expression levels of SOX-9 and Sirt1 proteins in TMJ tissue of the rats were significantly decreased(P<0.05);compared with model group,the expression levels of SOX-9 and Sirt1 proteins in TMJ tissue of the rats in resveratrol group were significantly increased(P<0.05),whereas the expression levels of MMP-13,PI3K,p-Akt,and p-mTOR proteins were significantly decreased(P<0.05).The RT-qPCR results showed that compared with control group,the expression levels of MMP-13,PI3K,Akt,and mTOR mRNA in TMJ tissue of the rats in model group were significantly increased(P<0.05),while the expression levels of SOX-9 and Sirt1 mRNA were significantly decreased(P<0.05);compared with model group,the expression levels of SOX-9 and Sirt1 mRNA in TMJ tissue of the rats in resveratrol group were significantly increased(P<0.05),whereas the expression levels of MMP-13,PI3K,Akt,and mTOR mRNA were significantly decreased(P<0.05).Conclusion:Resveratrol has therapeutic effect on TMJOA,and its mechanism may be related to the activation of Sirt1 and inhibition of the PI3K-Akt-mTOR signaling pathway.
3.Latent profile analysis of frailty among family caregivers of patients with dementia
Rui LI ; Yan SHI ; Xiao SUN ; Li WANG ; Guomin BU ; Jingyi WANG ; Xiaoqian KONG ; Wenjing XIA
Chinese Journal of Nursing 2024;59(16):2021-2029
Objective To explore the current status and potential subtypes of frailty among family caregivers of patients with dementia,and to analyze the related influencing factors of different subtypes.Methods Dementia patients and their family caregivers in 8 community health service centers in Shanghai from June to October 2023 were recruited by convenience sampling.General information questionnaire,Tilburg Frailty Indicator(TFI),Pittsburgh Sleep Quality Index(PSQI),Self-Rating Depression Scale(SDS),Zarit Caregiver Burden Interview(ZBI),and Connor-Davidson Resilience Scale(CD-RISC)were conducted for investigation.Latent profile analysis was used to explore the potential subtypes of frailty among family caregivers of patients with dementia.The influencing factors associated with the potential subtypes were identified by univariate analysis and multivariate Logistic regression analysis.Results A total of 470 family caregivers of patients with dementia were surveyed,and 46.17%of them suffered from frailty.Frailty among family caregivers of patients with dementia can be classified into 3 potential subtypes:comprehensive-low frailty subtype(70.64%),psychosocial-medium frailty subtype(19.57%),and physical-high frailty subtype(9.79%).Family caregivers of patients with dementia who had poor sleep quality and suffered from 2 or more chronic diseases were more likely to be classified into the physical-high frailty subtype(P<0.05).Family caregivers of patients with dementia who had higher levels of depression,lower mastery levels of caregiving knowledge and skills and spousal caregivers were more likely to be classified into the psychosocial-medium frailty subtype(P<0.05).Family caregivers of patients with dementia who had higher levels of resilience were more likely to be classified into the comprehensive-low frailty subtype(P<0.05).Conclusion The incidence of frailty among family caregivers of patients with dementia is at a high level with significant heterogeneity.It is suggested that medical staff should pay attention to the frailty of family caregivers,with a focus on family caregivers in the psychosocial-medium frailty subtype or physical-high frailty subtype,and take timely and targeted interventions according to the characteristics and influencing factors of different subtypes,so as to prevent or delay the occurrence and development of frailty.
4.Expert consensus on perioperative nursing management of nutrition for elderly patients with hip fractures (version 2023)
Chunhua DENG ; Xiaohua CHEN ; Zhihua YIN ; Yao JIANG ; Xiaoju TAN ; Yaping CHEN ; Junqin DING ; Luo FAN ; Leling FENG ; Yuyun GAN ; Xiaoyan GAO ; Jinli GUO ; Jing HU ; Chen HUANG ; Guiling HUANG ; Tianwen HUANG ; Yingchun HUANG ; Hui JIN ; Yan JIN ; Fangfang LI ; Hui LI ; Hui LIU ; Ping LIU ; Ning NING ; Lingyun SHI ; Guomin SONG ; Yani SUN ; Guangling WANG ; Jie WANG ; Qi WANG ; Xia WANG ; Xiaoyun WANG ; Yi WANG ; Songmei WU ; Jian YANG ; Yumei ZHANG ; Yang ZHOU ; Xiaoyan WANG ; Yuan GAO
Chinese Journal of Trauma 2023;39(5):394-403
Hip fractures are among the most common fractures in the elderly, presenting to be a leading cause of disability and mortality. Surgical treatment is currently the main treatment method for hip fractures. The incidence of perioperative malnutrition is increased after hip fractures in the elderly due to the comorbidities, decreased basal metabolic rate, accelerated protein breakdown, weakened anabolism and surgical stress. However, malnutrition not only increases the incidence of postoperative complications, but also leads to increased mortality, indicating an important role of perioperative nursing management of nutrition for the elderly patients with hip fractures. At present, there still lacks scientific guidance and application standards on perioperative nursing management of nutrition for the elderly patients with hip fractures. Therefore, the Orthopedic Nursing Committee of Chinese Nursing Association and the Editorial Board of Chinese Journal of Trauma organized relevant experts to formulate the Expert consensus on perioperative nursing management of nutrition for elderly patients with hip fractures ( version 2023) according to evidence-based medical evidences and their clinical experiences. Fourteen recommendations were made from aspects of nutrition screening, nutrition assessment, nutrition diagnosis, nutrition intervention and nutrition monitoring to provide guidance for perioperative nursing management of nutrition in elderly patients with hip fractures.
5.Correlation study between changes in intestinal microflora structure and immune indexes in newly treated patients with pulmonary tuberculosis
Jinhui XIE ; Rong YU ; Guomin SHI ; Xiaohua MA ; Sifang XIAO ; Yihang YI ; Ting ZHOU ; Yangen XIANG
Chinese Journal of Preventive Medicine 2021;55(12):1486-1490
To explore the correlation between the changes of the intestinal flora of newly treated pulmonary tuberculosis patients and the immune indicators of the body, and to provide a reference for the prevention and treatment of pulmonary tuberculosis. A single-center and case-control study was adopted. From October 2020 to April 2021, 43 patients with newly diagnosed tuberculosis in the Department of Tuberculosis, Affiliated Changsha Central Hospital,University of South China were selected as the control group. 43 cases of newly treated pulmonary tuberculosis (PTB), 43 healthy control (HC) during the same period, collected fresh feces and whole blood of subjects, and used Illumina Hiseq high-throughput sequencing technology to analyze 16S of all microorganisms in feces The V4 region of rRNA was amplified and sequenced, and the structure of the intestinal flora was analyzed by QIIME software. Use flow cytometry to determine the subject′s immune indicators (CD3 +, CD4 +, CD8 +, CD4 +CD25 +CD127 -Treg, CD14 +CD16 +, CD14 +CD16 -), and analyze the changes in intestinal flora and immune function in newly treated pulmonary tuberculosis patients Inherent connection. The χ2 test, t test, and Wilcox rank sum test were used to analyze the differences in age, gender, α diversity, and relative abundance of the two groups of people. Compared with the HC group, the alpha diversity of the intestinal flora in the PTB group decreased (shannon index: t=3.906, P=0.000 2; simpson index: Z=553, P=0.004 7; chao1 index: t=5.395, P=0.000 0). β diversity analysis showed that there were significant differences in the structure of the intestinal flora between the two groups ( P=0.000). Species difference analysis showed that at the phylum level, the relative abundance of Firmicutes in the PTB group was significantly lower than that in the HC group ( Z=486.0, P=0.000 5). At the genus level, there are 15 different bacterial genera between the two groups. In the PTB group, bifidobacterium, enterococcus, lactobacillus, anaerostipes, the relative abundance of the above 5 genera of veillonella is higher than that of the HC group ( P<0.05); Butyricimonas, clostridium, and broutella (blautia), coprococcus, dorea, lachnospira, roseburia, faecalibacterium, ruminococcus, the relative abundance of 10 bacterial genera including dialister was lower than that of the HC group ( P<0.05). Comparison of immune indexes between groups showed that CD14 +CD16 +monocytes (%) in the PTB group were higher than those in the HC group ( t=2.456, P=0.001 6<0.05), while CD14 +CD16 -monocytes (%) were lower than HC ( t=-4.368, P=0.000<0.05), while the differences in CD3 +, CD4 +, CD8 +, CD4 +/CD8 +and Treg (CD4 +CD25 +CD127 -) were not statistically significant ( P>0.05). Spearman correlation analysis showed that Firmicutes in the PTB group was negatively correlated with CD4 +/CD8 +, CD14 +CD16 +( r=-0.218, P=0.048; r=-0.245, P=0.025), and positively correlated with CD14 +CD16 -Correlation ( r=0.250, P=0.022); At the genus level, Faecalis is positively correlated with CD4 +/CD8 +and CD4 +( r=0.250, P=0.023; r=0.258, P=0.019); Rosella and CD3 +, CD8 +and CD14 +CD16 -are positively correlated ( r=0.27, P=0.024; r=0.219, P=0.046; r=0.027, P=0.039), and negatively correlated with CD14 +CD16 +( r=-0.280, P= 0.01). Changes in the structure of the intestinal flora of newly treated pulmonary tuberculosis patients may be one of the influencing factors of the immune function of the body. Targeted optimization of the structure of the intestinal flora and improvement of the body′s immunity may be used as an effective auxiliary treatment for pulmonary tuberculosis.
6.Correlation study between changes in intestinal microflora structure and immune indexes in newly treated patients with pulmonary tuberculosis
Jinhui XIE ; Rong YU ; Guomin SHI ; Xiaohua MA ; Sifang XIAO ; Yihang YI ; Ting ZHOU ; Yangen XIANG
Chinese Journal of Preventive Medicine 2021;55(12):1486-1490
To explore the correlation between the changes of the intestinal flora of newly treated pulmonary tuberculosis patients and the immune indicators of the body, and to provide a reference for the prevention and treatment of pulmonary tuberculosis. A single-center and case-control study was adopted. From October 2020 to April 2021, 43 patients with newly diagnosed tuberculosis in the Department of Tuberculosis, Affiliated Changsha Central Hospital,University of South China were selected as the control group. 43 cases of newly treated pulmonary tuberculosis (PTB), 43 healthy control (HC) during the same period, collected fresh feces and whole blood of subjects, and used Illumina Hiseq high-throughput sequencing technology to analyze 16S of all microorganisms in feces The V4 region of rRNA was amplified and sequenced, and the structure of the intestinal flora was analyzed by QIIME software. Use flow cytometry to determine the subject′s immune indicators (CD3 +, CD4 +, CD8 +, CD4 +CD25 +CD127 -Treg, CD14 +CD16 +, CD14 +CD16 -), and analyze the changes in intestinal flora and immune function in newly treated pulmonary tuberculosis patients Inherent connection. The χ2 test, t test, and Wilcox rank sum test were used to analyze the differences in age, gender, α diversity, and relative abundance of the two groups of people. Compared with the HC group, the alpha diversity of the intestinal flora in the PTB group decreased (shannon index: t=3.906, P=0.000 2; simpson index: Z=553, P=0.004 7; chao1 index: t=5.395, P=0.000 0). β diversity analysis showed that there were significant differences in the structure of the intestinal flora between the two groups ( P=0.000). Species difference analysis showed that at the phylum level, the relative abundance of Firmicutes in the PTB group was significantly lower than that in the HC group ( Z=486.0, P=0.000 5). At the genus level, there are 15 different bacterial genera between the two groups. In the PTB group, bifidobacterium, enterococcus, lactobacillus, anaerostipes, the relative abundance of the above 5 genera of veillonella is higher than that of the HC group ( P<0.05); Butyricimonas, clostridium, and broutella (blautia), coprococcus, dorea, lachnospira, roseburia, faecalibacterium, ruminococcus, the relative abundance of 10 bacterial genera including dialister was lower than that of the HC group ( P<0.05). Comparison of immune indexes between groups showed that CD14 +CD16 +monocytes (%) in the PTB group were higher than those in the HC group ( t=2.456, P=0.001 6<0.05), while CD14 +CD16 -monocytes (%) were lower than HC ( t=-4.368, P=0.000<0.05), while the differences in CD3 +, CD4 +, CD8 +, CD4 +/CD8 +and Treg (CD4 +CD25 +CD127 -) were not statistically significant ( P>0.05). Spearman correlation analysis showed that Firmicutes in the PTB group was negatively correlated with CD4 +/CD8 +, CD14 +CD16 +( r=-0.218, P=0.048; r=-0.245, P=0.025), and positively correlated with CD14 +CD16 -Correlation ( r=0.250, P=0.022); At the genus level, Faecalis is positively correlated with CD4 +/CD8 +and CD4 +( r=0.250, P=0.023; r=0.258, P=0.019); Rosella and CD3 +, CD8 +and CD14 +CD16 -are positively correlated ( r=0.27, P=0.024; r=0.219, P=0.046; r=0.027, P=0.039), and negatively correlated with CD14 +CD16 +( r=-0.280, P= 0.01). Changes in the structure of the intestinal flora of newly treated pulmonary tuberculosis patients may be one of the influencing factors of the immune function of the body. Targeted optimization of the structure of the intestinal flora and improvement of the body′s immunity may be used as an effective auxiliary treatment for pulmonary tuberculosis.
7.Spectrum change of rheumatic diseases in hospitalized children: a 12-year single-center experience of Shanghai
Tao ZHANG ; Haimei LIU ; Guomin LI ; Yu SHI ; Wen YAO ; Yifan LI ; Wanzhen GUAN ; Lijun ZHOU ; Fang LIU ; Hong XU ; Li SUN
Chinese Journal of Rheumatology 2020;24(2):120-124
Objective:To gain insight into the constitution of juvenile rheumatic diseases, treatment outcome and trends of rheumatic inpatients in past 12 years, and to improve awareness of juvenile rheumatic diseases.Methods:The clinical data of 5 950 patients in rheumatology department of the affiliated pediatric hospital of Fudan University (from 2005 to 2016) were analyzed retrospectively, and the chi-square test was used to compare and analyze the incidence.Results:Disease changes: ① The top three rheumatic diseases were Kawasaki disease (KD) (44.3%), Henoch-schoniein purpura (HSP) (35.4%), juvenile idiopathic arthritis (JIA)(9.6%). ② The number of all constitution of juvenile rheumatic diseases in hospital increased other than HSP. ③ The rheumatic diseases were increased from 17 to 37 kinds in the past 6 years. ④ The number of systemic lupus erythematosus (SLE) increased year by year (112/2 348 vs 197/3 602, χ2=1.41, P=0.235), as well as the severe SLE (35/112 vs 55/197, χ2=0.38, P=0.536). ⑤ The rate of rheumatic diseases complicated with macrophage activation (MAS) was 7.2‰(43/5 950). 12.9%(26/201) of systemic juvenile idiopathic arthritis(sJIA) were complicated with MAS, which was accounted for 60.5%(26/43) of total number of MAS in rheumatic diseases. In the last 6 years, there was a significant increase in the number of patients with MAS in patients with rheumatic diseases ( χ2=14.1, P<0.01) and sJIA( χ2=11.2, P<0.01). ⑥ 1.1%(64/5 950) of rheumatic diseases patients had lung lesions, juvenile dermatomyositis (JDM) accounted for 24.4%(20/82). In the last 6 years, the number of patients with lung lesions associated with rheumatic diseases increased significantly ( χ2=5.66, P=0.017). ⑦ The mortality rate of juvenile rheumatic diseases was only 3.7‰(22/5 950), and 45.5% occurred in SLE (10/22). The mortality rate of SLE decreased in last 6 years (5/112 vs 5/197, χ2=0.34, P=0.558). Conclusion:The constitution of juvenile rheumatic diseases in our center is decreasing for systemic vasculitis (KD, HSP), JIA, SLE, JDM in last 6 years. The annual total number of patients is relatively stable. But rare, difficult and critically illed cases increase year by year. Although SLE is still the primary cause of death in juvenile rheumatic diseases in recent 6 years, the mortality rate has decreased year by year.
8.Study on clinical separation characteristics of L-form of mycobacterium tuberculosis
Dongbin LIU ; Yangen XIANG ; Xiaohua MA ; Jianhua PAN ; Guomin SHI ; Yongjun CHEN ; Xuefang PENG
Journal of Chinese Physician 2019;21(3):351-355
Objective To explore the laboratory culture and identification of Mycobacterium tuberculosis L forms (MTB-L),isolation rate and drug resistance in smear-positive tuberculosis patients,and to improve clinical attention to MTB-L.Methods 222 smear-positive pulmonary tuberculosis patients treated in our hospital from September 2017 to December 2017 were randomly selected for MGIT 960 and 92-3TB-L liquid culture.After MGIT 960 was reported positive,acid-fast staining was performed on the precipitated smears of 92-3TB-L liquid medium for preliminary screening.The suspected L-positive strain culture was transformed into improved TSA-L solid medium to observe the colony characteristics and microscopic characteristics.The properties of the strain were confirmed by acid-fast staining and tuberculosis DNA amplification.Drug susceptibility and mutation sites of drug resistance genes were analyzed in MTB-L.Results Ⅰ-dentification of MTB-L:after the positive strain has been cultured,the colonies have the characteristics of "fried egg sample","particle-like" and " filament-like".MTB confirmed by tubercle DNA amplification experiments.Isolation rate:after cultured by MGIT 960 and Modified 92-3TB-L medium,the positive rate of single bacterial type was 50.90%,(113/222) the positive rate of both bacterial type and L type was 15.32% (34/222),and the positive rate of MTB-L type was 2.25% (5/222).Drug resistance:MTB-L was resistant to Streptomycin,Isoniazid,Rifampin,and Ethanol butylamine.No mutation was found in the drug resistance gene loci.Conclusions Clinical laboratory should routinely develop the culture of L-form of Mycobacterium tuberculosis bacteria,and increase the clinical attention to MTB-L.
9.The effect of artesunate on the expression of HO-1 induced by ESAT-6/CFP-10 of mycobacterium tuberculosis
Xiaohua MA ; Yangen XIANG ; Dongbin LIU ; Jianhua PAN ; Renhua FAN ; Rong YU ; Guomin SHI
Journal of Chinese Physician 2018;20(11):1647-1650
Objective To investigate the effect of artesunate (ASN) on the expression of Heme oxygenase-1 (HO-1) in THP-1 cells induced by the early secretory antigenic target-6 (ESAT-6) and culture filtrate protein-10 (CFP-10) antigens of Mycobacterium tuberculosis and to investigate its possible mechanism.Methods THP-1 ceils were cultured in vitro.The effects of ESAT-6 and CFP-10 on cell viability were detected by methyl thiazolyl tetrazolium (MTT) assay.THP-1 cells were pre-treated with or without ASN prior to incubation with or without ESAT-6 and CFP-10,the mRNA expression of HO-1 was detected by real time quantitative polymerase chain reaction (RT-qPCR) and Toll-like receptor 2 (TLR2) level was measured by Western blot.Results MTF assay showed that ESAT-6 and CFP-10 were non-toxic to cells in the range of 0-5 μg/ml.Compared with the control group,5 μg/ml ESAT-6 and 5 μg/ml CFP-10 could significantly increased the mRNA expression of HO-1 (P < 0.05).In addition,20 μg/ml ASN could significantly enhance the mRNA expression of HO-1 induced by ESAT-6 and CFP-10,and inhibit the expression of TLR2 induced by ESAT-6.Conclusions ASN in combination with ESAT-6 or CFP-10,may have potential value in treatment of pathogen-associated inflammatory diseases.
10.Long term effect of tocilizumab on refractory systemic juvenile idiopathic arthritis
Wen YAO ; Li SUN ; Haimei LIU ; Yu SHI ; Guomin LI ; Lijun ZHOU ; Hong XU
Journal of Clinical Pediatrics 2017;35(6):454-457
Objective To summarize the treatment experience of refractory systemic juvenile idiopathic arthritis (JIA) by tocilizumab, and to explore the cost-effective treatment. Methods The clinical data of 6 pediatric patients with refractory systemic JIA treated by tocilizumab from 2014 to June 2016 were retrospectively analyzed in the aspects of course and effectiveness of tocilizumab, steroid reduction, adverse reaction, and growth. Results The median age of the six patients (3 males and 3 females) was 6 years, and the course of disease were from 16 to 63 months. All patients were treated by other immunosuppressive agents or biological agents in addition to steroid and traditional anti-rheumatic drug therapy. The courses of tocilizumab treatment were from 7 to 26 months and the median time was 9.5 months. All 6 patients responded to tocilizumab and achieved the clinical remission at different time. After the induced remission, the interval of the treatment intervention was increased from 2 weeks up to 4 weeks in 3 cases, and no disease activity was observed. Except one case, another 5 cases reduced and stopped the use of hormones at 5.8 months after tocilizumab treatment. After hormones was reduced and discontinued, the growth was improved. All 6 patients had no serious adverse reactions. Conclusions Tocilizumab is safe and effective for patients with refractory JIAs. The steroid can be reduced in short time to improve growth. After remission is induced, the interval of the treatment intervention could be prolonged.

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