Objective To explore the expression of miR-21 and miR-19a in juvenile idiopathic arthritis (JIA) and the relationship among the key target genes (SOCS3,STAT3) in JAK/STAT pathways.Methods The venous blood from 33 cases of active JIA in Guangzhou Women and Children Medical Center were collected.All cases were divided into two groups:the systemic group (n=20),polyarthritis group (n=13).Twenty subjects were used as the normal control group.Peripheral blood mononuclear cells (PBMCs) were extracted and separated with Ficoll.miRNA was extracted and purified and real-time quantitative polymerase chain reaction (RT-PCR) was used to obtain cDNA.Target genes of miRNA were detected through Targetscan and RNA22.U6 was used for reference of miR-19a,miR-21 and β-actin were used for STAT3,SOCS3,IL-6,TNF-α mRNA.All the expression were detected by fluorescence quantitative PCR among the groups and calculated the result in standardized 2-ΔΔCT value,non-parametric test was used to test the differences.Results The expression of miR-21 were significantly reduced in the case group than the control group (Z=2.11,P=0.036),in which miR-21 was 7(7-8.5) times reduced than the SJIA group,6.49 (6-7) times than the pJIA group,the difference was statistically significant (Z=2.615,P=0.014 9;Z=2.654,P=0.0291).But no significant difference of miR-21 expression could be found between the SJIA and PJIA groups (Z=0.221,P =0.827 1).The expression of miR-19a was significantly reduced in the case group than the control group (Z=2.41,P=0.014),in which miR-19a was 11.3 (10-12.1) times to the SJIA group,12.2 (12-13.5) times to the pJIA group,the difference was statistically significant (Z=2.334,P=0.015 7;Z=2.414,P=0.026 6).But no significant difference could be detected in the miR-21 expression between the SJIA and the PJIA groups (Z=0.538,P=0.596).Software estimated that STAT3,SOCS3,TNF-α were the target genes of miR-21 and miR-19a in the JAK/STAT pathways respectively.Fluorescence quantitative PCR had shown that mRNA expression of STAT3 [6.24(2.81,7.54) and 3.97(1.81,5.75),P=0.001,0.008],TNF-α [3.03(2.07,3.80) and 3.42(2.46,4.68),P=0.002,0.001],IL-6[4.75(3.59,6.32) and 3.52(2.31,7.51),P=0.006,0.036],SOCS3[2.54(1.77,4.00) and 3.57(1.95,3.83),P=0.003,0.001] was higher in the case Group (SJIA group,the PJIA group) than the control group;STAT3 mRNA expression was negatively correlated with the miR-21 (r=-0.585 4,P=0.006 7;r=-0.613 4,P=0.044 7) and there was statistically significant difference.TNF-α,SOCS3 mRNA expression in the case group (SJIA group,PJIA group) was negatively correlated with the miR-19a.TNF-α (r=-0.664 2,P=0.001 4),SOCS3 (r=-0.790 3,P=0.000 1) of the SJIA group,was higher than those of the PJIA group TNF-α (r=-0.626 1,P=0.039 3),SOCS3 (r=-0.8824,P=0.003),the difference was significant.Conclusion The expression of miR-21,miR-19a in PBMC in the JIA patients are lower than the control group.The high expression of the target genes,miR-21,miR-19a of STAT3,SOCS3,TNF-α suggest that these genes might associate with,activating of JAK/STAT pathway.

Objective To investigate the effects of interleukin?17A on kidney injury induced by paraquat(PQ). Methods Seventy?two ICR mice were randomly divided into 3 groups:NS,PQ,and PQ+Ab (n=24 for each). The PQ?poisoning model was established by administering a gavage of PQ solution;mice in the PQ+Ab group were then administereda dose of anti?IL?17A antibody 2 hours later by i.p. injection,whereas the NS group were administered a corresponding volume of normal saline instead.The mice were killed at 8,24,48,or 72 h to obtain renal tissues and serum. An enzyme?linked immunosorbent assay(ELISA)was used to determine serum IL?17A,serum creatinine(SCr),and blood urea nitrogen (BUN)levels.Chemical colorimetry was used to detect the viability of myeloperoxidase(MPO )in renal tissue,and hematoxylin?eosin(HE)stain?ing was used to observe the renal pathologic changes. Immunohistochemistry(IHC)and PCR were used to examine IL?17A expression in renal tis?sues. Results Serum IL?17A,renal tissue MPO viabilities,BUN,and SCr were increased in the PQ and PQ+Ab groups,compared to those in the NS group(P<0.01). However,the above?mentioned parameters were lower in the PQ+Ab group than in the PQ group(P<0.01). Conclusion IL?17A promotes mouse kidney injury induced by acute PQ?intoxication through activating and/or recruiting neutrophils;therefore,blockade IL?17A,with antibody can attenuate the injury.

Objective To investigate the role of interleukin-17A (IL-17A) in acute paraquat (PQ)-induced lung injury in mice.Methods A total of 120 healthy SPF grade ICR male mice were randomly (random number) divided into three groups (n =40 in each):normal saline control group (NS),PQ poisoning group (PQ) and antibody neutralization group (PQ + Ab).Mice of PQ group and PQ + Ab group were given 5 mg/mL PQ by one gavage in a dosage of 25 mg/kg body weight,and 5 μg IL-17A neutralizing antibody intra-peritoneally administered immediately after PQ poisoning in PQ + Ab group;Equivalent volume of normal saline instead of PQ was given to mice of NS group.Six survival mice from each group were taken for experiment at 8 h,1 d,3 d,5 d,7 d after PQ poisoning:Wet to dry ratio (W/D) of lung was determined in mice of each group.HE staining of lung tissue was used to observe the histopathological changes under the light microscope and the pathological scores were graded;Serum interleukin-17A (IL-17A),interleukin-22 (IL-22),interleukin-6 (IL-6),transforming growth factor-β (TGF-β) were detected with enzyme linked immunosorbent assay (ELISA);Expression of interleukin-23 receptor (IL-23R) in lung tissue was determined with immunohistochemical;real-time fluorescence quantification PCR (qRT-PCR) was used to detect the expression of retinoic acid related solitary nuclear receptors' mRNA in lung tissue.Results After administration of PQ,W/D ratio increased (P ＜ 0.01),lung injury was observed in mice of PQ and PQ + Ab groups,levels of cytokines (IL-17A,IL-22,IL-6 and TGF-β) in serum elevated (P ＜0.05),and the expressions of IL-23R mRNA and RORγt mRNA increased (P＜0.01).But in PQ +Ab group,W/D ratio decreased (P ＜0.05),lung injury was alleviated,the levels of cytokines (IL-17A,IL-22,IL-6 and TGF-β) decreased (P ＜ 0.05),and the expressions of IL-23R mRNA and RORγt mRNA reduced (P ＜ 0.05).Conclusions Since IL-17A involves in the lung injury of the mice induced by acute paraquat poisoning,blockade of IL-17A significantly alleviates the acute lung injury in mice.

OBJECTIVE: To determine the effect of different tidal volume(Vt) ventilation on apoptosis and the expression of Bax and Bcl-2 in rat lungs. METHODS: Twenty-four healthy SD rats were randomly divided into 4 groups: a control group, a low Vt ventilation group (LV), a middle Vt ventilation group (MV), and a high Vt ventilation group (HV). Rats were subjected to different tidal volumes (10,20,and 40 mL/kg) for 2 h except the control group, which kept their own breath. We determined the lung histopathology score, W/D ratio and WBC in bronchoalceolar lavage fluid (BALF) to evaluate the lung injury and examine the apoptotic cell death, Bax and Bcl-2 protein expression by using TUNEL technique and immunohistochemistry 24 h after the operation. RESULTS: Compared with the control group, MV and HV increased lung histopathology score, W/D ratio, WBC in BALF, apoptosis index (AI )and Bax protein expression, but decreased Bcl-2 protein expression (P<0.05). These changes showed no significant difference between the control group and the low Vt ventilation group (P>0.05). CONCLUSION Low Vt ventilation contributed little to apoptosis. Higher Vt ventilation can improve Bax while inhibit Bcl-2 expression to aggravate apoptosis in rat lungs.
Animals ; Apoptosis ; physiology ; Female ; Lung ; pathology ; Male ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Respiration, Artificial ; adverse effects ; methods ; Tidal Volume ; physiology ; bcl-2-Associated X Protein ; metabolism

The non-neuronal cholinergic system, widely exists in prokaryotic, eukarytic, and even plant cells, however, it has not been investigated in preadipocytes and adipocytes. To search for evidence its existence in preadipocytes and adipocytes, the nicotinic acetylcholine receptor (nAChR) α7 subunit, acetyicholinesterase (ACHE) and choline acetyltransferase (CHAT) in 3T3-L1 cells were examined using immunohistochemical staining and Western blotting. The choline-regulated visfatin expression in 3T3-L1 preadipocytes was also tested by reverse transcriptase-PCR. Incubation with methyilycaconitine (10-6 to 10-4mol/L) for 12, 24 and 36 hours dose-dependently increased visfatin expression from 1.3- to 1.55-folds (P <0.01) with maximal induction at 24 hours with 10-4mol/L methyllycaconitine. Nicotine treatments (10-6 to 10-4 mol/L) for 12, 24 and 36 hours decreased visfatin expression; choline chloride (10-4 mol/L))suppressed visfatin expression in 3T3-L1 preadipocytes at 36 hours by 1.64 to 2.03 fold (P < 0.05) which was more effective as compared with nicotine. It was concluded that α7 nAChR was expressed in 3T3-L1 preadipocytes and involved in visfatin expression.

Summary: In order to confirm whether the mRNA levels of adiponectin in adipose tissue and mRNA levels of AdipoR1 in the skeletal muscles were correlated with the serum parameters of glucose and lipid metabolism and to clarify the regulation of adiponectin receptor gene expression in diabetic states, serum adiponectin, mRNA levels of adiponectin in adipose tissue and mRNA levels of AdipoR1 in the skeletal muscles were examined in type 2 diabetic rats. The model of type 2 diabetes was prepared by feeding high fat diet and injecting low dosage of streptozotocin (STZ). The diabetic rats were screened out by oral glucose tolerance test. One group of type 2 diabetic rats received rosiglitazone. The serum adiponectin concentration was detected by using ELISA and mRNA levels were examined by RT-PCR. The serum adiponectin levels and mRNA levels of adiponectin in adipose tissue of type 2 diabetic rats were significantly decreased as compared with the normal control rats (P<0.05, P<0.01 respectively). No siglificant changes were observed in the expression of adiponectin receptor 1 in the skeletal muscle of type 2 diabetic rats. The mRNA levels of adiponectin in adipose tissue were reversely correlated with serum insulin (r=-0.66, P<0.05), triglyceride (r=-0.58, P<0.05), cholesterol (r=-0.49, P<0.05), interleukin-6 (r=-0.49, P<0.05) and tumor necrosis factor (r=-0.43, P<0.05). The expression of adiponectin receptors was not altered in the skeletal muscle of Type 2 diabetic rats. The decreased serum adiponectin was caused by the decreased expression of adiponectin mRNA in adipose tissue rather than the adiponectin receptors in the skeletal muscle, which could be improved by rosiglitazone.

Objective To evaluate the prognosis of patients with sepsis in the emergency department using the modified CURB-65 score.Methods We retrospectively analyzed the clinical data of 143 patients with sepsis who were first diagnosed at the emergency department of the First Hospital of China Medical University (between January 2014 and January 2015),assessed their CURB-65 and sequential organ failure assessment (SOFA) scores,and modified the CURB-65 scoring system by adding some indexes of the prognosis of sepsis.We analyzed the prognostic value of each scoring systems in the diagnosis of sepsis using the receiver-operating characteristic curve.Results The modified CURB-65,CURB-65,and SOFA scores had independent abilities for early prediction of the prognosis of sepsis.The area under the curve and the Youden index of the modified CURB-65 score were highest,which are superior to the traditional CURB-65 and SOFA scores.Conclusion The modified CURB-65 score can predict the prognosis of sepsis in its early stage.In addition,the assessment method is simple and convenient;hence,it is useful for assessing the condition of patients with sepsis and providing an early treatment.

The present study evaluated the application of three dimensional echocardigraphy (3DE) in the diagnosis of atrial septal defect (ASD) and the measurement of its size by 3DE and compared the size with surgical findings. Two-dimensional and real-time three dimensional echocardiography (RT3DE) was performed in 26 patients with atrial septal defect, and the echocardiographic data were compared with the surgical findings. Significant correlation was found between defect diameter by RT3DE and that measured during surgery (r=0.77, P<0.001). The defect area changed significantly during cardiac cycle. Percentage change in defect size during cardiac cycle ranged from 6%-70%. Our study showed that the size and morphology of atrial septal defect obtained with RT3DE correlate well with surgical findings. Therefore, RT3DE is a feasible and accurate non-invasive imaging tool for assessment of atrial septal size and dynamic changes.
Echocardiography, Three-Dimensional ; Heart Septal Defects, Atrial/*diagnosis ; Heart Septal Defects, Atrial/*pathology ; Heart Septal Defects, Atrial/surgery ; Young Adult

Objective To explore the expression of inflammasomes (NLRP3,NLRP12) and related signal proteins in the peripheral blood mononuclear cells (PBMCs) of patients with juvenile idiopathic arthritis (JIA).Methods Samples of children with definite diagnosis of active JIA in Guangzhou Women and Childrens' Medical Center were collected retrospectively.Fifty-five cases were included,among whom 30 were systemic type and 25 were joint type.Blood samples of 22 healthy controls were collected at the same time.Peripheral blood single nuclear cell (PBMCs) were separated and DNA were extracted and reverse transcription (RT) to cDNA.Fluorescent quantitative polymerase chain reaction (PCR) was used to detect NLRP3,NLRP12,ASC,and capase-1 in groups and the difference in their expression between groups were analyzed.Enzyme linked immunosorbent assay (ELISA) was utilized to test plasma levels of interleukin (IL)-6 IL-1,IL-4,IL-10,and their correlation were analyzed.Results The expression of NLRP3,NLRP12,ASC,Capase-1 in the case group (general-group and joint-group) were higher than those in the control group (P＜0.05),but there was no significant difference in the expression levels between groups (P＞0.05).The IL-1 concentration of the case group (body-type group,joint-group) was higher than the control group (P=0.001,U=l) (P=0.001,U=14),however,the level of IL-4 of the case group (body-type group,joint-group) was not significantly different from the control group (U=662,P=0.13) (U=823,P=0.535),IL-I0 of the systemic group was higher than that of the control group (U=750,P=0.023),while there was no difference between groups (U=672,P=0.212).There were no significant difference in the levels of IL-1 (U=658,P=0.408),IL-4 (U=475,P=0.068),IL-10 (U=475,P=0.195) between groups.The NLRP3 mRNA relative expression levels of the case group and the ASC (r=0.44,P=0.013 4) was significant,in addition,IL-1 (P=0.001,R=0.58),erythrocyte sedimentation rate (ESR) (r=0.415,P=0.039),C reactive protein (CRP) (r=0.438,P=0.046) were positively correlated with NLRP12 relative mRNA expression level and ASC (r=0.583 7,P=0.007),CRP (r=0.46,P=0.031 6),ESR (r=0.003,P=0.56),CD8+ T (r=0.414,P=0.036).Conclusion The abnormal expression of JIA inflammasomes in peripheral blood mononuclear cells (NLRP3,NLRP12) may be associated with juvenile idiopathic arthritis.

Objective Myeloid-derived suppressor cell (MDSC) have the potential to suppress autoimmune T cell response, while the levels and function of it had not been studied in juvenile idiopathic arthritis (JIA). Therefore, we used flow cytometry to detect MDSC in peripheral blood of JIA and analyzed its correlation with cytokines, providing clinical basis for the generation and function of MDSCs in JIA. Methods A total of 34 patients with JIA were included. The disease activity was evaluated by calculating the 28-joint Disease Activity Score (DAS28)based on clinical information. JIA patients were divided into the active disease group and inactive disease group according to the DAS28 score. Twenty healthy controls were recruited from the Health Care Section of our hospital.The frequency of MDSC in peripheral blood from JIA patients and healthy controls were determinedby flow cytometry; enzyme-linked immunosorbent assay (ELISA) was used to detect plasma Interleukin (IL)-1β, IL-6, IL-10 and IL-17A of JIA patients and healthy controls; RT-quantitative polymerase chain reaction (qPCR) was performed to detect the expression levels of signal transducer and activator of transcription 3 (STAT3),IL-17A gene in patients with JIA and control subjects,and Spearman correlation was used to investigate the association between MDSC and DAS28 score, cytokines;non-parametric test and Spearman test were used for two group comparisons and correlation analysis, respectively.Results The frequency of MDSC [(6.2±4.2)% vs (1.2±1.6)%, Z=-5.258, P<0.01], and the levels of plasma cytokines IL-1β [(175±306) pg/ml vs (66±29) pg/ml, Z=-2.246, P=0.034], IL-6 [(86.6±257.2) pg/ml vs (14.0± 2.6)pg/ml,Z=-3.123,P=0.002),IL-10[(44±36)pg/ml vs(24±6)pg/ml,Z=-2.166,P=0.03],IL-17A[(9.1±17.6) pg/ml vs (2.7±0.4) pg/ml, Z=-3.965, P<0.01] incre-ased significantly in JIA patients compared with healthy control group, while STAT3 had no differences (P>0.05). The frequency of MDSC correlated positively with DAS28 and erythrocyte sedimentation rate(ESR) disease activity (r=0.447, P=0.037; r=0.456, P=0.033), IL-1β (r=0.496, P=0.044), IL-10 (r=0.498, P=0.035); negatively cor-related with the STAT3 mRNA (r=-0.522, P=0.041); but not correlated with the IL-6, IL-17A by Spearman's test. Conclusion We have found that the frequency of MDSC in JIA increases and is associated disease activity, closely associated with inflammatory cytokines. The frequency of MDSC can be used as a clinical indicator to reflect inflammatory activity in JIA, However,the current knowledge of MDSC is incomplete and further experiment is required.