1.Activity of Codonopsis canescens against rheumatoid arthritis based on TLRs/MAPKs/NF-κB signaling pathways and its mechanism.
Yu-Jie WANG ; Xiao-Yu ZHONG ; Xin-Hong WANG ; Yuan-Han ZHONG ; Lin LIU ; Fang-Yuan LIU ; Jin-Xiang ZENG ; Ji-Xiao ZHU ; Xiao-Lang DU ; Min LI ; Gang REN ; Guo-Yue ZHONG ; Xiao-Min WANG
China Journal of Chinese Materia Medica 2022;47(22):6164-6174
		                        		
		                        			
		                        			This paper aims to explore the activity of Codonopsis canescens extract against rheumatoid arthritis(RA) based on the Toll-like receptors(TLRs)/mitogen-activated protein kinases(MAPKs)/nuclear factor kappa B(NF-κB) signaling pathways and its mechanism. The ultra-performance liquid chromatography-quadrupole time-of-flight/mass spectrometry(UPLC-Q-TOF-MS) was used to identify the components of C. canescens extract. Forty-eight male SD rats were randomly divided into six groups, namely the normal group, the model group, the methotrexate(MTX) tablet group, and the low, medium, and high-dose C. canescens extract(ZDS-L, ZDS-M, and ZDS-H) groups, with 8 rats in each group. The model of collagen-induced arthritis in rats was induced by injection of bovine type Ⅱ collagen emulsion. MTX(2.5 mg·kg~(-1)), ZDS-L, ZDS-M, and ZDS-H(0.3 g·kg~(-1), 0.6 g·kg~(-1), and 1.2 g·kg~(-1)) were administrated by gavage. Rats in the normal group and the model group received distilled water. MTX was given once every three days for 28 days, and the rest medicines were given once daily for 28 days. Body weight, degree of foot swelling, arthritis index, immune organ index, synovial histopathological changes, and serum levels of tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), and interleukin-6(IL-6) were observed. Protein expressions of TLR2, TLR4, NF-κB p65, p38 MAPK, and p-p38 MAPK in rats were determined by Western blot. Thirty-four main components were identified by UPLC-Q-TOF-MS, including 15 flavonoids, 7 phenylpropanoids, 4 terpenoids, 4 organic acids, 2 esters, and 2 polyalkynes. As compared with the normal group, the body weight of the model group was significantly decreased(P<0.01), and foot swelling(P<0.05, P<0.01), arthritis index(P<0.01), and the immune organ index(P<0.01) were significantly increased. The synovial histopathological injury was obviously observed in the model group. The serum levels of inflammatory factors TNF-α, IL-1β, and IL-6 were significantly increased(P<0.01), and the protein expression levels of TLR2, TLR4, NF-κB p65, p-p38 MAPK/p38 MAPK in the synovial tissue were significantly increased(P<0.01) in the model group. As compared with the model group, the body weights of the ZDS dose groups were increased(P<0.01), and the degree of foot swelling(P<0.01) and the arthritis index were decreased(P<0.05, P<0.01). The immune organ index was decreased(P<0.01) in the ZDS dose groups, and the synovial tissue hyperplasia and inflammatory cell infiltration were alleviated. The serum levels of TNF-α, IL-1β, and IL-6 were significantly decreased(P<0.05, P<0.01), and the protein expression levels of TLR2, TLR4, NF-κB p65, p-p38 MAPK/p38 MAPK were decreased(P<0.05, P<0.01) in the ZDS dose groups. C. canescens extract containing apigenin, tricin, chlorogenic acid, aesculin, ferulic acid, caffeic acid, and oleanolic acid has a good anti-RA effect, and the mechanism may be related to the inhibition of TLRs/MAPKs/NF-κB signaling pathways.
		                        		
		                        		
		                        		
		                        			Animals
		                        			;
		                        		
		                        			Cattle
		                        			;
		                        		
		                        			Male
		                        			;
		                        		
		                        			Rats
		                        			;
		                        		
		                        			Arthritis, Experimental/drug therapy*
		                        			;
		                        		
		                        			Arthritis, Rheumatoid/drug therapy*
		                        			;
		                        		
		                        			Body Weight
		                        			;
		                        		
		                        			Codonopsis/chemistry*
		                        			;
		                        		
		                        			Interleukin-6/blood*
		                        			;
		                        		
		                        			NF-kappa B/genetics*
		                        			;
		                        		
		                        			p38 Mitogen-Activated Protein Kinases/metabolism*
		                        			;
		                        		
		                        			Plant Extracts/therapeutic use*
		                        			;
		                        		
		                        			Rats, Sprague-Dawley
		                        			;
		                        		
		                        			Signal Transduction
		                        			;
		                        		
		                        			Toll-Like Receptor 2/metabolism*
		                        			;
		                        		
		                        			Toll-Like Receptor 4/metabolism*
		                        			;
		                        		
		                        			Tumor Necrosis Factor-alpha/pharmacology*
		                        			
		                        		
		                        	
2.Detection Value of EB Virus DNA, IL-2, and IL-6 Level in Peripheral Blood of Children with Infectious Mononucleosis.
Qiang ZHANG ; Qing-Feng FANG ; Zhi YANG ; Bi-Quan CHEN
Journal of Experimental Hematology 2022;30(4):1262-1266
		                        		
		                        			OBJECTIVE:
		                        			To investigative the detection value of EB virus DNA (EBV-DNA), interleukin-2 (IL-2), and interleukin-6 (IL-6) level in peripheral blood of children with infectious mononucleosis (IM).
		                        		
		                        			METHODS:
		                        			A total of 59 children clinically confirmed with IM in Anhui Provincial Children's Hospital from January 2018 to September 2020 were enrolled as IM group, while other 30 healthy children undergoing physical examination during the same period were enrolled as healthy group. The level of EBV-DNA load, IL-2, and IL-6 were compared between the two groups, and their diagnostic values for IM children were explored. According to the median level of EBV-DNA load, positive children were divided into high viral load group and low viral load group. The hepatomegaly and splenomegaly, and levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), IL-2, and IL-6 were compared between the two groups. The relationship between EBV-DNA load and IL-2, IL-6 levels were explored.
		                        		
		                        			RESULTS:
		                        			The positive rate of EBV-DNA was 67.80% in IM group, which was significantly higher than 10.00% in healthy group (P<0.001), and the levels of serum IL-2 and IL-6 were also significantly higher than healthy group (P<0.001). The results of ROC curve analysis showed that AUC of IL-2 combined with IL-6 and EBV-DNA load was 0.948, which was significantly greater than that of IL-2, IL-6, and EBV-DNA load alone (0.847, 0.728, 0.789) (P<0.001). The cut-off value of IL-2 and IL-6 was 15.545 pg/ml and 56.560 pg/ml, respectively. Both the proportions of cases with moderate to severe hepatomegaly and splenomegaly in high viral load group were significantly higher than those in low viral load group (P<0.01, P<0.05). The levels of ALT, AST, and IL-2 in high viral load group were significantly higher than those in low viral load group (P<0.001), as well as IL-6 (P<0.01). In high and low viral load groups, EBV-DNA load was positively correlated with IL-2 and IL-6 (in high viral load group, rIL-2=0.598, rIL-6=0.416; in low viral load group, rIL-2=0.621, rIL-6=0.527, P<0.001).
		                        		
		                        			CONCLUSION
		                        			The detection of EBV-DNA load combined with IL-2 and IL-6 can improve the diagnostic accuracy of IM, and EBV-DNA load, IL-2 and IL-6 levels are related to the disease progression.
		                        		
		                        		
		                        		
		                        			Child
		                        			;
		                        		
		                        			DNA, Viral
		                        			;
		                        		
		                        			Epstein-Barr Virus Infections
		                        			;
		                        		
		                        			Hepatomegaly
		                        			;
		                        		
		                        			Herpesvirus 4, Human
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Infectious Mononucleosis/diagnosis*
		                        			;
		                        		
		                        			Interleukin-2/blood*
		                        			;
		                        		
		                        			Interleukin-6/blood*
		                        			;
		                        		
		                        			Splenomegaly
		                        			
		                        		
		                        	
3.Serum interleukin-2 receptor α as a clinical biomarker in patients with systemic lupus erythematosus.
Jia Yi TIAN ; Xia ZHANG ; Gong CHENG ; Qing Hong LIU ; Shi Yang WANG ; Jing HE
Journal of Peking University(Health Sciences) 2021;53(6):1083-1087
		                        		
		                        			OBJECTIVE:
		                        			To investigate the clinical relevance of serum interleukin-2 receptor α (IL-2Rα) in patients with systemic lupus erythematosus (SLE).
		                        		
		                        			METHODS:
		                        			One hundred and seven SLE patients and 39 healthy controls with comparable age and gender were recruited at Peking University People's Hospital from January 2019 to December 2020. Complete clinical data in 107 SLE patients at baseline and follow-up were collected. SLE disease activity index 2000 (SLEDAI-2K) was used to assess the disease activity of the SLE patients. The serum level of IL-2Rα in the SLE patients and healthy controls was measured using enzyme-linked immunosorbent assay (ELISA). The association between serum IL-2Rα and clinical and laboratory parameters was investigated. Mann-Whitney U test or t test, Chi-square test and Spearman correlation were used for statistical analysis.
		                        		
		                        			RESULTS:
		                        			The serum IL-2Rα levels were significantly higher in the SLE patients [830.82 (104.2-8 940.48) ng/L], compared with those in the healthy controls [505.1 (78.65-1 711.52) ng/L] (P < 0.001). Association analysis showed that the increased serum IL-2Rα was positively associated with SLEDAI-2K scores and anti-nucleosome antibody (r=0.357, P < 0.001; r=0.25, P=0.027, respectively). Thirty-six of 107 (33.6%) SLE patients had lupus nephritis. Serum IL-2Rα levels were significantly higher in the patients accompanied with lupus nephritis [1 102.14 (126.52-8 940.48) ng/L] than in the patients without lupus nephritis [743.89 (104.19-4 872.06) ng/L] (P=0.032). The patients in the high IL-2Rα group had more lupus nephritis compared with those in the low IL-2Rα group (40.8% vs. 19.4%, P=0.031). Meanwhile, SLEDAI-2K scores were found significantly higher in the high IL-2Rα group than in the low IL-2Rα group [10 (3-21) vs. 7 (3-16), P=0.001]. With the improvement of disease activity in the SLE patients after conventional treatments, serum levels of IL-2Rα [1 119.1 (372.25-2 608.86) ng/L] in the week 12 decreased significantly compared with the baseline [1 556.73 (373.08-8 940.48) ng/L] (P=0.042).
		                        		
		                        			CONCLUSION
		                        			Serum IL-2Rα may be used as a biomarker of disease activity in patients with SLE. There is certain correlation between serum IL-2Rα and renal involvement in SLE.
		                        		
		                        		
		                        		
		                        			Biomarkers/blood*
		                        			;
		                        		
		                        			Case-Control Studies
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Interleukin-2 Receptor alpha Subunit/blood*
		                        			;
		                        		
		                        			Lupus Erythematosus, Systemic/diagnosis*
		                        			
		                        		
		                        	
4.PF-04620110, a Potent Antidiabetic Agent, Suppresses Fatty Acid-Induced NLRP3 Inflammasome Activation in Macrophages
Seung Il JO ; Jung Hwan BAE ; Seong Jin KIM ; Jong Min LEE ; Ji Hun JEONG ; Jong Seok MOON
Diabetes & Metabolism Journal 2019;43(5):683-699
		                        		
		                        			
		                        			BACKGROUND: Chronic inflammation has been linked to insulin resistance and type 2 diabetes mellitus (T2DM). High-fat diet (HFD)-derived fatty acid is associated with the activation of chronic inflammation in T2DM. PF-04620110, which is currently in phase 1 clinical trials as a selective acyl-CoA:diacylglycerol acyltransferase-1 (DGAT1) inhibitor, is a potent anti-diabetic agent that may be important for the regulation of chronic inflammation in T2DM. However, the mechanisms by which PF-04620110 regulates fatty acid-induced chronic inflammation remain unclear. METHODS: PF-04620110 was used in vitro and in vivo. DGAT1-targeting gRNAs were used for deletion of mouse DGAT1 via CRISPR ribonucleoprotein (RNP) system. The activation of NLRP3 inflammasome was measured by immunoblot or cytokine analysis in vitro and in vivo. RESULTS: Here we show that PF-04620110 suppressed fatty acid-induced nucleotide-binding domain, leucine-rich-repeat-containing receptor (NLR), pyrin-domain-containing 3 (NLRP3) inflammasome activation in macrophages. In contrast, PF-04620110 did not change the activation of the NLR family, CARD-domain-containing 4 (NLRC4), or the absent in melanoma 2 (AIM2) inflammasomes. Moreover, PF-04620110 inhibited K⁺ efflux and the NLRP3 inflammasome complex formation, which are required for NLRP3 inflammasome activation. PF-04620110 reduced the production of interleukin 1β (IL-1β) and IL-18 and blood glucose levels in the plasma of mice fed HFD. Furthermore, genetic inhibition of DGAT1 suppressed fatty acid-induced NLRP3 inflammasome activation. CONCLUSION: Our results suggest that PF-04620110 suppresses fatty acid-induced NLRP3 inflammasome activation.
		                        		
		                        		
		                        		
		                        			Animals
		                        			;
		                        		
		                        			Blood Glucose
		                        			;
		                        		
		                        			Clinical Trials, Phase I as Topic
		                        			;
		                        		
		                        			Clustered Regularly Interspaced Short Palindromic Repeats
		                        			;
		                        		
		                        			Diabetes Mellitus, Type 2
		                        			;
		                        		
		                        			Diacylglycerol O-Acyltransferase
		                        			;
		                        		
		                        			Diet, High-Fat
		                        			;
		                        		
		                        			Fatty Acids
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			In Vitro Techniques
		                        			;
		                        		
		                        			Inflammasomes
		                        			;
		                        		
		                        			Inflammation
		                        			;
		                        		
		                        			Insulin Resistance
		                        			;
		                        		
		                        			Interleukin-18
		                        			;
		                        		
		                        			Interleukins
		                        			;
		                        		
		                        			Macrophages
		                        			;
		                        		
		                        			Melanoma
		                        			;
		                        		
		                        			Mice
		                        			;
		                        		
		                        			Plasma
		                        			;
		                        		
		                        			Ribonucleoproteins
		                        			;
		                        		
		                        			RNA, Guide
		                        			
		                        		
		                        	
5.Clinical significance of detection of soluble interleukin 2 receptor alpha chain in the assessment of rheumatoid arthritis disease activity.
Jia Jia XU ; Yan WANG ; He SUN ; Ru Lin JIA ; Xue Wu ZHANG ; Yang MENG ; Li Li REN ; Xiao Lin SUN
Journal of Peking University(Health Sciences) 2018;50(6):975-980
		                        		
		                        			OBJECTIVE:
		                        			To evaluate soluble interleukin-2 receptor alpha chain (sIL-2Rα, sCD25) in serum for the determination of rheumatoid arthritis (RA) activity.
		                        		
		                        			METHODS:
		                        			Peripheral blood was collected from 108 patients with RA, 39 patients with osteoarthritis (OA) and 50 healthy control subjects, and synovial fluids were from 40 patients with RA. The sera from the patients with RA, the disease control group (osteoarthritis), the healthy control group, and the synovial fluids of the RA patients were detected by enzyme-linked immunosorbent assay (ELISA).The clinical manifestations and laboratory parameters of the patients with RA were recorded and the correlation with the serum sCD25 level was analyzed.
		                        		
		                        			RESULTS:
		                        			The serum sCD25 concentration in RA group was (2 886±1 333) ng/L, the serum sCD25 concentration in OA group was (2 090±718) ng/L, and the serum sCD25 concentration in healthy group was (1 768±753) ng/L. The serum sCD25 level in the patients with RA was significantly higher than that in the disease controls and healthy controls (P<0.001). Sensitivity of serum sCD25 in the diagnosis of RA was 66.1% and specificity was 83.0%;serum sCD25 levels and erythrocyte sedimentation rate (r=0.321, P=0.001), C-reactive protein (r=0.446, P<0.001), DAS28 score (r=0.324, P<0.001), joint tenderness count (r=0.203, P=0.024), D-dimer levels (r=0.383, P<0.001), age (r=0.24, P=0.007), IgG (r=0.207, P=0.028), HRF-IgG (r=0.345, P=0.034) showed a significant positive correlation, and disease duration (r=-0.206, P=0.021) showed a negative correlation with sCD25;In patients with rheumatoid arthritis, the positive rates of serum ESR, CRP, and sCD25 were 14.3% (2 cases), 14.3% (2 cases), and 71.4% (10 cases) in the low disease activity group. The positive rates of serum ESR, CRP and sCD25 in the moderate disease activity group were 94.2% (49 cases), 82.7% (43 cases), and 86.5% (45 cases). The positive rates of serum ESR, CRP, and sCD25 in the high disease activity group were 100% (42 cases), 95.2% (40 cases), and 90.5% (38 cases);36 cases of ESR and/or CRP were negative (about 33.3%) in 108 patients, serum sCD5 levels of 17 cases in these 36 cases (about 47.2%)increased, of which 14 cases (about 82.4%) had a DAS28 score higher than 3.2.
		                        		
		                        			CONCLUSION
		                        			The serum sCD25 has a high specificity for diagnosis of RA and a poor sensitivity. The serum level is closely related to the activity of RA, indicating that sCD25 may be involved in the inflammatory process of RA and may become a new inflammatory marker of RA. It is more meaningful for detection of serum sCD25 when RA is active, but ESR and/or CRP is negative.
		                        		
		                        		
		                        		
		                        			Arthritis, Rheumatoid/pathology*
		                        			;
		                        		
		                        			Biomarkers/analysis*
		                        			;
		                        		
		                        			Blood Sedimentation
		                        			;
		                        		
		                        			C-Reactive Protein
		                        			;
		                        		
		                        			Case-Control Studies
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Interleukin-2 Receptor alpha Subunit/analysis*
		                        			;
		                        		
		                        			Osteoarthritis
		                        			;
		                        		
		                        			Synovial Fluid/chemistry*
		                        			
		                        		
		                        	
6.Angiotensin-(1-9) ameliorates pulmonary arterial hypertension via angiotensin type II receptor.
Seung Ah CHA ; Byung Mun PARK ; Suhn Hee KIM
The Korean Journal of Physiology and Pharmacology 2018;22(4):447-456
		                        		
		                        			
		                        			Angiotensin-(1-9) [Ang-(1-9)], generated from Ang I by Ang II converting enzyme 2, has been reported to have protective effects on cardiac and vascular remodeling. However, there is no report about the effect of Ang-(1-9) on pulmonary hypertension. The aim of the present study is to investigate whether Ang-(1-9) improves pulmonary vascular remodeling in monocrotaline (MCT)-induced pulmonary hypertensive rats. Sprague-Dawley rats received Ang-(1-9) (576 µg/kg/day) or saline via osmotic mini-pumps for 3 weeks. Three days after implantation of osmotic mini-pumps, 50 mg/kg MCT or vehicle were subcutaneously injected. MCT caused increases in right ventricular weight and systolic pressure, which were reduced by co-administration of Ang-(1-9). Ang-(1-9) also attenuated endothelial damage and medial hypertrophy of pulmonary arterioles as well as pulmonary fibrosis induced by MCT. The protective effects of Ang-(1-9) against pulmonary hypertension were inhibited by Ang type 2 receptor (AT₂R) blocker, but not by Mas receptor blocker. Additionally, the levels of LDH and inflammatory cytokines, such as TNF-α, MCP-1, IL-1β, and IL-6, in plasma were lower in Ang-(1-9) co-treated MCT group than in vehicle-treated MCT group. Changes in expressions of apoptosis-related proteins such as Bax, Bcl-2, Caspase-3 and -9 in the lung tissue of MCT rats were attenuated by the treatment with Ang-(1-9). These results indicate that Ang-(1-9) improves MCT-induced pulmonary hypertension by decreasing apoptosis and inflammatory reaction via AT₂R.
		                        		
		                        		
		                        		
		                        			Angiotensins*
		                        			;
		                        		
		                        			Animals
		                        			;
		                        		
		                        			Apoptosis
		                        			;
		                        		
		                        			Arterioles
		                        			;
		                        		
		                        			Blood Pressure
		                        			;
		                        		
		                        			Caspase 3
		                        			;
		                        		
		                        			Cytokines
		                        			;
		                        		
		                        			Hypertension*
		                        			;
		                        		
		                        			Hypertension, Pulmonary
		                        			;
		                        		
		                        			Hypertrophy
		                        			;
		                        		
		                        			Interleukin-6
		                        			;
		                        		
		                        			Lung
		                        			;
		                        		
		                        			Monocrotaline
		                        			;
		                        		
		                        			Plasma
		                        			;
		                        		
		                        			Pulmonary Fibrosis
		                        			;
		                        		
		                        			Rats
		                        			;
		                        		
		                        			Rats, Sprague-Dawley
		                        			;
		                        		
		                        			Receptor, Angiotensin, Type 2
		                        			;
		                        		
		                        			Vascular Remodeling
		                        			
		                        		
		                        	
7.Relationship between expression of peripheral blood HLA-DR, CD4CD25 regulatory T cells, IL-17 and IL-27 with liver damage in children with human cytomegalovrius infection.
Li-Li ZHU ; Ling XU ; Jun WANG
Chinese Journal of Contemporary Pediatrics 2018;20(7):554-558
OBJECTIVETo study the relationship between the expression of peripheral blood HLA-DR, CD4CD25 regulatory T cells, IL-17 and IL-27 with liver damage in children with human cytomegalovirus (HCMV) infection.
METHODSTwenty-one HCMV children with liver damage and twenty-one HCMV children without liver damage were enrolled in this study. The expression of peripheral blood HLA-DR and CD4CD25 regulatory T cells was detected by flow cytometry. Plasma levels of IL-17 and IL-27 were measured using ELISA.
RESULTSThe plasma levels of IL-17 and IL-27 in children with liver damage were significantly higher than in those without liver damage, while the expression of peripheral blood CD4CD25 regulatory T cells was lower than in those without liver damage (P<0.05). Plasma IL-17 and IL-27 levels were negatively correlated with the expression of peripheral blood CD4CD25 regulatory T cells (P<0.01).
CONCLUSIONSImmune imbalance mediated by CD4CD25 regulatory T cells and over-expression of IL-17 and IL-27 may be involved in the pathogenesis of liver damage in children with HCMV infection.
CD4 Antigens ; immunology ; Cytomegalovirus ; physiology ; Cytomegalovirus Infections ; blood ; complications ; genetics ; Female ; Flow Cytometry ; HLA-DR Antigens ; genetics ; immunology ; Humans ; Infant ; Interleukin-17 ; blood ; genetics ; Interleukin-2 Receptor alpha Subunit ; immunology ; Interleukins ; blood ; genetics ; Liver ; injuries ; metabolism ; Liver Diseases ; blood ; etiology ; immunology ; Male ; T-Lymphocytes, Regulatory ; immunology
8.Clinical assessment after human adipose stem cell transplantation into dogs
Seok Hee LEE ; Erif M N SETYAWAN ; Yoo Bin CHOI ; Jeong Chan RA ; Sung Keun KANG ; Byeong Chun LEE ; Geon A KIM
Journal of Veterinary Science 2018;19(3):452-461
		                        		
		                        			
		                        			Adipose tissue-derived stem cell (ASCs) are an attractive source of stem cells with therapeutic applicability in various fields for regenerating damaged tissues because of their stemness characteristics. However, little has reported on evaluating adverse responses caused by human ASC therapy. Therefore, in the present study, a clinical assessment after human ASC transplantation into dogs was undertaken. A total of 12 healthy male dogs were selected and divided into four groups: saline infusion, saline bolus, ASC infusion, and ASC bolus groups. Physical assessment and blood analysis were performed following ASC transplantation, and the concentrations of angiogenic factors, and pro- and anti-inflammatory cytokines were measured by enzyme-linked immunosorbent assay (ELISA). There were no adverse vital sign responses among the dogs. Blood analyses revealed no remarkable complete blood count or serum chemistry results. ELISA results for angiogenic and anti-inflammatory factors including matrix metalloproteinase 9 (MMP9), vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), hepatocyte growth factor (HGF), and interleukin-10 (IL-10) were significantly higher in the two ASCs groups than in the controls. In conclusion, this study demonstrated that transplantation of human ASCs produced no adverse effects and could be used safely in dogs. In addition, human ASCs could be involved in modulating secretions of angiogenic factors including MMP9, VEGF, bFGF, and HGF and anti-inflammatory factor IL-10.
		                        		
		                        		
		                        		
		                        			Angiogenesis Inducing Agents
		                        			;
		                        		
		                        			Animals
		                        			;
		                        		
		                        			Blood Cell Count
		                        			;
		                        		
		                        			Chemistry
		                        			;
		                        		
		                        			Cytokines
		                        			;
		                        		
		                        			Dogs
		                        			;
		                        		
		                        			Enzyme-Linked Immunosorbent Assay
		                        			;
		                        		
		                        			Fibroblast Growth Factor 2
		                        			;
		                        		
		                        			Hepatocyte Growth Factor
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Interleukin-10
		                        			;
		                        		
		                        			Male
		                        			;
		                        		
		                        			Matrix Metalloproteinase 9
		                        			;
		                        		
		                        			Stem Cell Transplantation
		                        			;
		                        		
		                        			Stem Cells
		                        			;
		                        		
		                        			Transplantation
		                        			;
		                        		
		                        			Vascular Endothelial Growth Factor A
		                        			;
		                        		
		                        			Vital Signs
		                        			
		                        		
		                        	
9.Expression of Serum HMGB-1 in Patients with Secondary Hemo-phagocytic Lymphohistiocytosis and Its Clinical Significance.
Dong-Jiao WANG ; Ju-Juan WANG ; Meng SONG ; Xin GAO ; Li-Min DUAN ; Tian TIAN ; Hong-Xia QIU
Journal of Experimental Hematology 2016;24(3):878-883
OBJECTIVETo investigate the expression levels and clinical significance of serum high mobility group box 1 (HMGB-1) in patients with secondary hemophagocytic lymphohistiocytosis (sHLH).
METHODSSerum HMGB-1 levels were determined by using enzyme linked immunosorbent assays (ELISA) in 51 sHLH patients and 15 healthy contrlols. Other laboratory data, including soluble interleukin-2 receptor (sCD25), white blood cells (WBC), hemoglobin (Hb), platelet (Plt), fibrinogen (FIB), lactate dehydrogenase (LDH), triglyceride (TG), alanine transaminase (ALT), aspartate aminotransferase (AST), serum ferritin (SF), C reactive protein (CRP), and blood sedimentation rate (ESR) were also collected.
RESULTSSerum HMGB-1 levels in the newly diagnosed group were significantly higher than that in the control group (P<0.01). Serum HMGB-1 levels in the newly diagnosed lymphoma-associated HLH (LHLH) group were significantly higher than that in non-HLH group, including infection-associated HLH (IHLH) and autoimmune-associated HLH (AHLH) group (P<0.05); The serum HMGB-1 levels in the clinical remission group were significantly lower than that in the newly diagnosed group (P<0.05), however, serum HMGB-1 was not decreased significantly in the progression/relapsed group, compared with the newly diagnosed group (P>0.05). Serum HMGB-1 levels in newly diagnosed sHLH patients positively correlated with sCD25 (r=0.62, P<0.01) and ESR (r=0.55, P<0.05). The receiver operating characteristic curves (ROC) for serum HMGB-1 levels of sHLH patients and healthy controls produced a cutoff value at 15.3 µg/L, with its 90% sensitivity and 99% specificity, respectively. In addition, an optimal cutoff value for HMGB-1 was 27.4 µg/L in the patients LHLH and non-HLH (AHLH+IHLH) with 96% sensitivity and 81% specificity, separately.
CONCLUSIONSerum HMGB-1 levels possesses an important clinically significance for disease diagnosis, differential diagnosis, evaluation of nosographic activity and treatment efficacy in the patients with sHLH.
C-Reactive Protein ; analysis ; Case-Control Studies ; Diagnosis, Differential ; Enzyme-Linked Immunosorbent Assay ; Fibrinogen ; analysis ; HMGB1 Protein ; blood ; Humans ; Interleukin-2 Receptor alpha Subunit ; blood ; L-Lactate Dehydrogenase ; blood ; Leukocytes ; Lymphohistiocytosis, Hemophagocytic ; blood ; Lymphoma ; Sensitivity and Specificity ; Treatment Outcome
10.Effects of ulinastatin on immune function of patients with severe burn injury.
Juncong LI ; Chao HU ; Hongming YANG ; Yongming YAO
Chinese Journal of Burns 2016;32(6):345-350
OBJECTIVETo primarily evaluate the effects of ulinastatin on immune function of patients with severe burn injury.
METHODSForty patients with severe burn admitted to our ward from March 2013 to October 2015, conforming to the study criteria, were divided into conventional treatment group (CT, n=20) and ulinastatin treatment group (UT, n=20) according to the random number table and patient's consent. After admission, patients in group CT received antishock treatment, antibiotic treatment, debridement, skin grafting, and nutrition support, etc. On the basis of the above-mentioned treatment, patients in group UT received intravenous drip of ulinastatin from first day after admission twice a day, with a dosage of 8×10(5) U every time, for 7 days in addition. Peripheral venous blood samples were collected from patients in groups CT and UT on post treatment day (PTD) 1, 3, 5 and 7, respectively. Twenty healthy volunteer were selected as health control group (HC), and peripheral venous blood samples were collected on the first day of the study. Percentage of CD4(+) CD25(+) regulatory T lymphocytes (Tregs) was determined by flow cytometer. The proliferative activity of T lymphocytes was detected by microplate reader (denoted as absorbance value). Content of interleukin 2 (IL-2) in culture supernatant of T lymphocytes, and content of IL-4 and γ interferon (IFN-γ) in serum were detected by enzyme-linked immunosorbent assay. Expression of human leukocyte antigen-DR (HLA-DR) on CD14(+) monocytes was determined by flow cytometer. Data were processed with analysis of variance for repeated measurement, chi-square test, and LSD-t test.
RESULTS(1) Compared with that of volunteer in group HC, the percentage of CD4(+) CD25(+) Tregs of patients in group CT was significantly increased from PTD 1 to 7 (with t values from 13.303 to 26.043, P values below 0.01). Compared with that in group CT, the percentage of CD4(+) CD25(+) Tregs of patients in group UT was significantly decreased on PTD 5 and 7 (with t values respectively 8.317 and 15.071, P values below 0.01). (2) The proliferative activity of T lymphocytes of patients in group CT on PTD 1, 3, 5, and 7 was respectively 0.71±0.11, 0.61±0.15, 0.54±0.12, and 0.67±0.17, which was significantly lower than that in group HC (1.21±0.22, with t values from 8.686 to 11.957, P values below 0.01). The proliferative activity of T lymphocytes of patients in group UT on PTD 3, 5, and 7 were respectively 0.81±0.11, 0.85±0.14, and 1.08±0.13, which was significantly higher than that in group CT (with t values from 4.808 to 8.568, P values below 0.01). (3) Compared with those of volunteer in group HC, content of IL-2 in culture supernatant of T lymphocytes of patients in group CT was significantly decreased from PTD 1 to 7 (with t values from 8.073 to 9.288, P values below 0.01), content of IL-4 in serum of patients in group CT was significantly increased from PTD 1 to 7 (with t values from 18.926 to 41.451, P values below 0.01), and content of IFN-γ in serum of patients in group CT was significantly decreased from PTD 1 to 7 (with t values from 4.543 to 27.659, P values below 0.01). Compared with those in group CT, content of IL-2 in culture supernatant of T lymphocytes of patients in group UT was significantly increased from PTD 3 to 7 (with t values from 6.507 to 8.869, P values below 0.01), content of IL-4 in serum of patients in group UT was significantly decreased from PTD 3 to 7 (with t values from 6.922 to 8.843, P values below 0.01), and content of IFN-γ in serum of patients in group UT was significantly increased on PTD 5 and 7 (with t values respectively 5.369 and 13.521, P values below 0.01). (4) The percentages of CD14(+) monocytes with positive expression of HLA-DR of patients in group CT on PTD 1, 3, 5, and 7 were respectively (28±6)%, (25±7)%, (25±7)%, and (39±10)%, which were significantly lower than the percentage of volunteer in group HC [(87±8)%, with t values from 16.323 to 25.645, P values below 0.01]. The percentages of CD14(+) monocytes with positive expression of HLA-DR of patients in group UT on PTD 3, 5, and 7 were respectively (40±6)%, (42±9)%, and (49±10)%, which were significantly higher than those in group CT (with t values from 3.071 to 7.324, P values below 0.01).
CONCLUSIONSOn the basis of CT, additional ulinastatin intervention can decrease CD4(+) CD25(+) Tregs percentage, improve the immune function of T lymphocytes and T helper cells, and increase expression of HLA-DR on CD14(+) monocytes of patients with severe burn injury, thus improve the immune function of patients.
Burns ; drug therapy ; immunology ; Cells, Cultured ; Debridement ; Enzyme-Linked Immunosorbent Assay ; Glycoproteins ; therapeutic use ; Humans ; Interferon-gamma ; blood ; Interleukin-2 ; metabolism ; Interleukin-4 ; blood ; Monocytes ; immunology ; Skin Transplantation ; T-Lymphocytes, Regulatory ; immunology
            
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