Objective To observe the developmental changes of projection and termination of proprioceptive afferent fibers in the mouse spinal cord. Methods Parvalbumin (PV) immunohistochemistry was used to label the proprioceptive afferents. Single and dual immunofluorescence histochemistry were used to examine the growth pattern of proprioceptive afferents and their relationships with motoneurons in the spinal ventral horn (VH). The stained sections were observed under a confocal laserscanning microscope. Results PV-like immunoreactive (LI) proprioceptive fibers first appeared in the dorsal column on embryonic (E) day 14, then entered the gray matter on El5 and reached the intermediate gray matter and VH more obviously on E16. The number and intensity of PV-LI proprioceptive afferent fibers and punctata increased in the VH with age and reached a maximum during earlier postnatal (P) period (P0-P7). After P14, the number and intensity of proprioceptive afferents gradually decreased. The proprioceptive terminals seemed to form close relationship with motoneurons from E17. Conclusion The present study indicates that the somatotopic organization of proprioceptive afferents in the spinal cord is established during the late embryonic and early postnatal stages. These results provide evidence for understanding the development of the reflex movements.

The present study was designed to examine the developmental changes in projection and termination of nociceptive and proprioceptive afferent fibers in the spinal cord by labeling those two fibers with calcitonion gene-related peptide (CGRP) and parvalbumin (PV)separately in mouse embryos and neonatal pups aged embryonic day 15 to posanatal day 3 (E15 -P3). CGRP-like immunoreactive (LI)nociceptive fibers first appeared in the superficial dorsal horn (DH) at E16. The afferent projections extended laterally to the DH and entered into the deep portions of the DH at E17 and E18. After birth, the projection pattern of CGRP-LI fibers remained unchanged but the intensity of afferent terminals increased in the superficial laminae and their branching patterns became more complicated. In addition,CGRP-LI collaterals that projected into the contralateral DH were also examined after E16. Around birth, the contralateral projections were also found originated from the lateral part of the DH. PV-LI proprioceptive afferents were first observed entering the gray matter at E15 and reached the intermediate gray matter (IG) and the ventral horn (VH) more obviously on E16. The number and intensity of PV-LI fibers increased in the the VH with age and reached a maximum during earlier postnatal period ( P0-P3 ). The proprioceptive terminals seemed to form close relationship with motoneurons in the VH from E17. Our results indicate that the somatotopic organization of nociceptive and proprioceptive afferents in the spinal cord both are established during the late embryonic and early postnatal stages. These results help to understand the development of the sensory transmission in more details.

objective To study the role of anti-EBV antibody in the development of systemic lupus erythematosus(SLE).Methods Peripheral blood was obtained from 55 SLE patients and 29 normal controls,and anti-EBV-VCA IgG,IgA,IgM antibody was detected by enzyme-linked immunosorbent assay[ELISA).Totat RNA was extracted from peripheral blood of 33 SLE patients and 26 normal controls and then reverse transcribed into complementary DNA.Real-time PCR technique was used to determine gene expressions at the transcription level.Comparison of anti-EBV antibody level and interferon inducible gene expression was made between SLE Datients and normal controls by t-test.The associations between anti-EBV antibody level,lupus activitv and IFN inducible gene expression were analyzed by Spearman's correlation analysis.Results (1)The levels of anti-EBV antibodies in SLE patients were significantly increased as compared to those in the normal controls[IgG(41±6)vs(19±6)U/ml,IgA(15.4±1.8)vs(8.3±2.1)U/ml,IgM(7.8±1.0)vs(3.7±1.2)U/ml]cantly elevated in SLE Datients as compared to normal controls[IFN scores 11±13 vs 0±3](all P<0.05),but not correlated with the levels of anti-EBV antibody in patients(all P>0.05).Conclusion Anti-EBV antibody is overproduced in SLE patients but not associated with disease activity as well as the expression level of several major types of I IFN inducible genes,suggesting that EBV infection may not be a major factor mediating the activation of IFN pathway and consequently the exacerbation of SLE.

Acute Kidney Injury ; etiology ; Hemolytic-Uremic Syndrome ; complications ; etiology ; therapy ; Humans ; Shiga Toxin ; toxicity

Objective To explore the expression levels of interferon-inducible genes in patients with systemic lupus erythematosus ( SLE) , and to validate these gene expressions as potential biomarkers for the differentiation of disease flare and infection.Methods Peripheral blood was obtained from 48 SLE, 16 rheumatoid arthritis ( RA) patients and 26 normal controls, and total RNA was extracted and reverse transcribed into complementary DNA.Real-time PCR technique was used to determine the gene expressions of MX1, OASL,OAS1, ISG15 and LY6E at transcription level.Univariate logistic regression analysis and multivariate conditional logistic regression model were applied to analyze 5 related factors for infection or activity.Results ( 1 ) The expression levels of MX1, OASL, 0AS1, ISG15, and LY6E mRNA in SLE patients were significantly increased as compared with normal controls ( P all < 0.01 ) , while the expression levels of OASL,OAS1 ,ISG15 and LY6E mRNA in SLE patients were also higher than those in RA patients (P all <0.05 ).(2)There were no significant difference between male and female patients of the 5 gene expression in SLE patients.(3) By logistic regression analysis, ISG15 and LY6E were independent risk factors for active SLE patients (P <0.01) , OASL expression was an independent risk factor for SLE patients with infection ( P = 0.003 ).Conclusion All the 5 interferon inducible genes are highly expressed in SLE patients, in which ISG15 and LY6E are independently associated with disease flare, while OASL may be helpful for the evaluation of infection in SLE patients.

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 in vivo effects of allogeneic mesenchymal stem cells transplantation (MSCT) on OAZ signaling pathway in patients with systemic lupus erythematosus (SLE).Methods Isolated and expand human MSCs from bone marrow cells or umbilical cord of healthy donors were infused into SLE patients. Peripheral blood cells were collected from 10 pre-MSCT patients as well as 1 week and 4 week post-MSCT, and RNA was extracted and reverse transcripted to cDNA. mRNA expression levels of OAZ and Id1-3 were measured by using real-time PCR. Serum levels of IL-10, IL-12, IL-21, CCL2 and ANA were tested by ELISA. Relationships of the gene expression levels with levels of cytokines and ANA were analyzed. Results mRNA expression levels of OAZ, Id1 and Id3 gene in patients with SLE were significantly decreased at week 1(△C:12.4±1.1, 9.7±1.9, 9.7±1.9, 2.1±1.0) and at week 4 (△Ct:13.3±1.2, 10.4±1.5,10.8±1.2, 2.1±1.2) after MSCT when compared to those of the pre-MSCT (△Ct:11.0±0.9, 7.4±2.1, 7.8±2.1, 0.1±1.5 respectively, P all＜0.05). Levels of IL-10, IL-21 and ANA were significantly lower 4 week after MSCT than those before (P＜0.05); while level of CCL2 was higher than pre-MSCT (P＜0.05). Cytokines and ANA levels 1 week after MSCT were not differentially changed comparing to those of the pre-MSCT. Alteration of OAZ mRNA expression levels pre- and post-MSCT were negatively correlated with changes of ANA, IL-21levels and positively correlated with changes of IL-12/IL-10 and CCL2 levels. Conclusions The expression of genes involving in the OAZ signaling pathway is effectively reduced along with the alteration of several cytokines and ANA after allogeneic MSCT in SLE patients. OAZ signaling pathway may play an important role in MSCT treatment for SLE.

Carbon Monoxide ; analysis ; Spectroscopy, Fourier Transform Infrared ; methods

Objective To study the role of OIf-1/EBF associated zinc finger protein (OAZ),a transcription factor encoded by a positional systemic lupus erythematosus (SLE) candidate gene,in the function of mesenchymal stem cells (MSC) of SLE patients by silencing this gene.Methods OAZ mRNA levels of bone marrow MSC obtained from 5 SLE patients and 5 healthy controls were detected by real-time PCR.Bone marrow MSC obtained from 6 SLE patients were incubated with specific siRNAs for 3 days,then cells were harvested for OAZ measurement,Idl-3 and CCL2 mRNA levels were tested by real-time PCR,and levels of CCL2 were detected in culture supernatants using ELISA.Differences between groups were analyzed using t-test or MannWhitney test.Results ① OAZ mRNA levels of bone marrow MSC were significantly elevated in SLE patients (0.013±0.016) compared to healthy controls (0.001±0.000,P=0.009).② After OAZ silencing,the expression levels of OAZ,Id1,Id2 and ld3 mRNA were significantly decreased (△Ct 10.3±0.7,15.2±1.6,8.1±1.4,10.5±0.6 vs 8.7±0.7,14.1±1.2,7.1±1.5,9.8±0.6) (P all ＜0.05).③ Both the expression levels of CCL2 mRNA (△Ct 2.2±1.1 vs 3.0±1.1 ) and the levels of CCL2 protein in culture supernatants [(341±29) pg/ml vs (304±19) pg/ml] were significantly increased in OAZ silencing group comparing to those in the control group (P all ＜0.05).Conclusion OAZ gene expression is significantly elevated in bone marrow MSC of SLE patients.OAZ may affect autoantibody production in SLE patients by regulating CCL2 expression.

Objective To analyze the impacts of early enteral nutrition (EN) and parenteral nutrition (PN) on the prognosis of acute pancreatitis.Method By searching relevant literature between January 1996 and August 2013 in Chinese and English databases including Wanfang,VIP,CNKI,PubMed,the Cochrane Library,CBM,and EMABSE,we compared the prognosis of acute pancreatitis after EN or PN in terms of casefatality rate,infections,multiple organ failure,and other complications.Result Compared with PN,early EN significantly reduced the case-fatality rate [OR =0.37,95％ CI (0.23,0.58),P ＜ 0.000 1],decreased the infection rate [OR =0.24,95％ CI (0.15,0.39),P ＜0.000 01],shortened hospital stay [MD =-9.87,95％ CI (-10.84,-8.89),P ＜ 0.000 01],and reduced complications [OR =0.26,95％ CI (0.12,0.58),P =0.001 0] in patients with acute pancreatitis,although the incident of multi-organ failure showed no significant difference [OR =0.35,95％ CI (0.10,1.19),P =0.09].Conclusions For patients with acute pancreatitis,early EN is superior than PN in terms of case-fatality rate,infection rate,hospital stay,and complications.Therefore,it should be applied in such patients whenever condition allows.