Objective To investigate the regulatory effects of umbilical cord-derived mesenchymal stem cells (UC -MSCs) on Th17 cells and related cytokines in patients with systemic lupus erythematosus (SLE). Methods Human UC-MSCs were isolated and expanded and infused into fourteen SLE patients. Clinical changes were evaluated before and after transplantation by SLE disease activity index (SLEDAI), 24-hour urine protein, serum albumin and complement C3. The percentages of CD3 +CD8-IL17A + T cells in peripheral blood were detected by flow cytometry. Concentrations of plasma IL-6, TGF-β, IL-17A, IL-22were determined by enzyme-linked immunosorbent assay (ELISA). UC-MSCs and peripheral blood mononindependent samples t-test. Results SLEDAI scores decreased significantly at 3 month (7.8±1.2, t=2.19) and 6 month (6.9±0.9, t=4.2) after UC-MSCs transplantation than pre-transplantation level (10.4±0.9, P< 0.05). Twenty-four-hour proteinuria decreased significantly 6 months after MSCs infusion [(1489±260) mg vs (2454±322) mg, t=2.6, P<0.05]. Meanwhile, serum albumin and complement C3 levels had increased significantly since 1 month after transplantation (P<0.05). The percentages of peripheral blood CD3+CD8-IL17A+T cells decreased obviously in 1 week, 1 month and 6 months after UC-MSCs transplantation (all P<0.05). The coculture of UC-MSCs with PBMC from SLE patients resulted in a statistically significant reduction of CD3+CD8-IL17A+T cells percentage in PBMC (P<0.05), but was not in a dose dependent manner. No change of plasma IL+6, TGF-β, IL-17A and IL-22 levels was observed after UC-MSCs transplantation (P>0.05).Conclusion UC-MSCs transplantation down-regulates the percentages of CD3+CD8-IL17A+T cells in SLE patients, which may be one of the mechanisms for its therapeutic effect in refractory SLE.

Objective To study the role of the expression levels of 5 type Ⅰ interferon (IFN)-inducible genes (LY6E, OAS1, OASL, MX1, and ISG15) in the diagnosis and disease activity evaluation of systemic lupus erythematosus (SLE). Methods Peripheral blood was obtained from 68 SLE patients, 50 patients with other connective tissue diseases and 26 normal controls, and total RNA was extracted and reverse transcribed into complementary DNA. Real-time PCR technique was used to determine gene expressions at transcription level. An IFN score for each individual was calculated according to the expression of 5 1FN genes. Comparisons of gene expression and IFN score were made among groups. The genes expression levels in patients with SLE were analyzed using receiver operative characteristic curve. The association between IFN scores and disease activity, as assessed by the SLEDAI scores and 24 h proteinuria, was analyzed using Spearman correlation analyses. Results ① The expression levels of MX1, OASL, OAS1, ISG15 and LY6E mRNA in SLE patients were significantly increased as compared with normal controls and disease controls (P all＜0.01 ).② IFN scores in SLE patients (17.9±29.1) were significantly increased as compared with normal controls (0±3.3)and disease controls (3.0±8.1) (P all＜0.01 ). ③ IFN scores area under the ROC curve (AUCROC) was 0.846. When The IFN scores reached 2.56, its sensitivity and specificity for the diagnosis of SLE were 93.1%and 78.3%, respectively. ④ Levels of IFN score was positively correlated with SLEDAI scores (r=0.256,P＜0.05) and 24 h proteinuria (r=0.337, P＜0.05). Conclusion The 5 IFN-inducible genes are highly expressed in SLE patients. IFN score level is valuable for the diagnosis of SLE and a high IFN score is usually associated with an elevated disease activity.

Objective To investigate the apoptosis of bone marrow mesenchymal stem cells(BMSCs)from systemic lupus erythematosus(SLE)patients and the expression of apoptotic molecules.Methods BMSCs were isolated from bone marrow of SLE patients and normal controls by density eentrifugation and adhesive culture in vitro.The apoptosis of BMSCs were evaluated by TUNEL assay.The expressions of Fas.Bcl-2 and the activity of Caspase 8 were detected by flow cytometry.Real-time PCR technique was used to determine the gene expressions of Fas,Bcl-2,Bax,Bcl-w,Caspase 8 and Apaf-1.Meanwhile,cytochrome C was detected by immunocytochemistry.Statistical analysis was conducted with t-test and Mann-Whitney rank test.Results The percentage of apoptotic BMSCs increased in SLE patients compared with healthy donors[(64±10)%vs [14±9)%,U=0,P<0.05 ].The expression of Bcl-2 in BMSCs of SLE patients was lower than the normal controls at protein leve[(11±9)%vs(56±18)%,U=0,P<0.05 ],and mRNA level(0.2±0.2vs 2.4±0.7,U=24,P<0.05).More cytochrome C positive pellets in the cytosolic fraction could be detected in BMSCs from SLE patients compared with healthy controls [(56±21)%vs (16±16)%,U=1,P<0.05].The activity of Caspase 8 was enhanced[(49±14)%vs(16±12)%,U=0.P<0.05 ],although with no significant difierence at mRNA Ievel.Both groups expressed Fas but with no significant difference (U=19,P>0.05).Conclusion BMSCs from SLE patients undergo more apoptosis,the mechanisms may be associated with the down regulation of Bcl-2,up-regulation of Cytoehrome C in cytoplasm and the activation of Caspase 8,which directs the intrinsic and extrinsic apoptosis pathways.

OBJECTIVE:To observe the curative effect and safety of compound glycyrrhizin injection in the treatment of neonatal hepatitis syndrome(NHS).METHODS:68 neonates with NHS were randomly divided into treatment and control gro_up(n=34),undertaking intravenous administration of glycyrrhizin injection and shengmai injection respectively,both at a dose of 3ml/(kg?d) for 2 weeks consecutively.RESULTS:The liver function in the treatment group after treatment was significantly better than that in the control group(P

Objective To explore the role of intracellular reactive oxygen species (ROS) in the senescence of bone marrow mesenchymal stem cells(BMSCs)in patients with systemic lupus erythematosus(SLE) and the underlying mechanisms that controls the intracellular ROS levels in vitro. Methods Human bone marrow aspirates were collected from iliac of eight donors and eight SLE patients and cultured in vitro.Morphological appearance of BMSCs at different passages was examined by inverted microscope. Nuclear size was measured by fluorescence microscope. BMSCs were monitored using the senescence associated β-galacto-sidase (SAβ-gal) assay to characterize senescence in vitro. The quantification of intracellular ROS production was detected by flow cytometry. Real-time PCR technique was used to determine the gene expressions of PI3K, KRas, NRas and FoxO3 at transcription level. The expression of FoxO3, phospho-FoxO3 (p-FoxO3),AKT and phospho-AKT (p-AKT) protein were determined by Western blot analysis. Statistical analysis was conducted with t-test and Mann-Whitney rank test.Results There were no differences in morphology and nuclear size[(31.4±4.5) vs (28.2±4.8) μm, P=0.628] of BMSCs between SLE patients and normal controls.The percentage of SA β-gal positive BMSCs from SLE patients was higher than that from healthy controls [(31.8±9.0)% vs (12.4±0.7)%, P＜0.05]. Intracellular ROS levels of BMSCs from SLE patients increased more significantly than healthy donors in vitro (34600±9600 vs 17 958±5400, P＜0.05). No significant differences in the expression of PI3K, NRas, KRas and FoxO3 from SLE subjects were observed at mRNA levels compared with normal controls, though all showed a similar upward trend. The expression of p-FoxO3 and p-AKT of BMSCs from SLE patients increased significantly compared with healthy controls at protein levels.Conclusion These data suggest that BMSCs from SLE patients aged more quickly, with high SA β-gal activity and up-regulation of intracellular ROS, which is associated with up-regulation of p-FoxO3 and pAKT at protein levels. These results indicate that bone marrow mesenchymal cell senescence may be associated with the pathogcnesis of SLE by maintaining the lifespan of BMSCs.

ObjectiveTo investigate the effect of IκB kinase (IKK-β) on migration and proliferation of bone marrow mesenchymal stem cells(BMSCs) from patients with systemic lupus erythematosus (SLE).MethodsHuman bone marrow aspirates were collected from iliac of six donors and six SLE patients and cultured in vitro.Migration of BMSCs were observed by wound healing and transwell migration assays.Proliferation of BMSCs was quantified by cell counting kit-8 assay.Total RNA was extracted and reverse transcribed into complementary DNA.Real-time PCR technique was used to determine the gene expression of IKK-β at transcription level.The expression of IKK-β and phospho-IKK-β(p-IKK-β) protein were determined by Western blotting analysis.Statistical analysis was conducted with or Mann-Whitney rank test.Results① The migration rate of BMSCs from SLE patients(5.2±3.8)‰ were significantly reduced as compared with normal controls (7.0±2.9)‰(P＜0.05 ).The proliferation of BMSCs of SLE patients (0.21±0.49)was lower than that from healthy controls ( 1.00±0.35 )(P＜0.05 ).②) No difference in IKK-β3 mRNA expression between SLE ( 1.9± 1.4) subjects and normal controls (1.9±2.4) (P＞0.05).IKK-β protein expression of BMSCs from SLE patients (1.41 ±0.19) increased significantly compared with healthy controls (0.93±1.24) (P＜0.05).③ Inhibitor of IKK-β caused a significant increase in cell migration (3.3±1.6)‰ and proliferation (1.13±0.26) of BMSCs from SLE patients compared with untreated cells (2.3±1.1)‰ and (0.81±0.17),respectively (P＜0.05).ConclusionMigration and proliferation of BMSCs are significantly decreased in SLE patients.IKK-β may be involved in migration and proliferation of BMSCs.

Objective To identify the association of HLA-DQA1 and-DQ B1 alleles with vitiligo in Chinese Hans in Anhui province.Methods Polymerase chain reaction sequence-specific primer (PCR-SSP) method was used to analyze the distribution of HLA-DQA1 and-DQB1 alleles among 187 patients with vitiligo and 273 healthy controls.Results The frequencies of HLA-DQA1*0302,-DQB1*0303,-DQB1*0503 alleles were significantly increased in the patients with vitiligo compared with those in the controls,and HLA-DQA1*0501 allele frequency was mark edly decreased.HLA-DQA1*0302,-DQA1*0601,-DQB1*0303,-DQB1*0503 alleles were p ositively associated,whereas HLA-DQA1*0501 allele was negatively associated wit h childhood vitiligo,and HLA-DQB1*0303 allele was positively associated with ad ult vitiligo,in comparison with those in the controls.The frequency of HLA-DQB 1*0303 allele was significantly increased in localized vitiligo patients compare d with that in the controls,whereas frequencies of HLA-DQA1*0302,-DQB1*0303,-DQB1*0503 alleles were significantly increased and the frequency of HLA-DQA1*050 1 allele was markedly decreased in generalized vitiligo patients.Conclusions HLA-DQA1*0302,-DQA1*0601,-DQB1*0303 and-DQB1*0503 alleles could be the suscep tible alleles of vitiligo,while HLA-DQA1*0501 allele could be the protective al lele in Chinese Hans in Anhui province.There may be different genetic backgroun ds between childhood and adult patients,and between localized and generalized vitiligo.