OBJECTIVE:To determine the plasma concentration of mycophenolic acid (MPA) in renal transplantation patients by HPLC-Fluoremetry. METHODS: The sample was subjected to precipitation of proteins using 5% Zinc Sulfate methanol saturated solution,and the supernatant (20 ?L) was taken for sample injection and determination on Zorbax Eclipse XDB C18 with mobile phase consisted of acetonitrile-methanol-0.2 mol?L-1 glycine buffer(18∶2∶80,pH=9.0) at a flow rate of 1.0 mL?min-1. The column temperature was of 25℃;the excitation wavelength(EX) was 342 nm and the emission wavelength(EM) was 425 nm. RESULTS: The linear range of MPA was 0.5～40 mg?L-1,with its lowest limit of quantitation at 0.5 mg?L-1. The methodology recovery was 98.23%～101.00%;the extraction recovery of MPA was 91.56%～94.46%;the intra-day RSD was 0.64%～3.22% and the inter-day RSD was 5.12%～6.10%. CONCLUSION: The method is sensitive,rapid,accurate,convenient,and applicable for the quantitative determination of plasma concentration of MPA in renal transplantation patients.

Acute Disease ; Atrial Fibrillation ; diagnosis ; etiology ; therapy ; Child ; Child, Preschool ; Electrocardiography ; Humans ; Infant ; Myocarditis ; complications ; virology ; Virus Diseases ; complications

Objective To explore skeletal‐related events (SREs) clinical factors and analysis prognosis factors on patients with non‐small cell lung cancer(NSCLC) with bone metastases .Methods We collected clinical data of pathology confirmed 383 patients with non‐small cell lung cancer between April 2007 and January 2007 in the third affiliated hospital of the third military medical uni‐versity .It was used to screening for Emission Computed Tomography (ECT ) for bone metastases .And then it was need to con‐firmed for CT ,MRI or PET‐CT or pathology .Statistics in patients between clinical features and the SREs prediction factor with Univariate and Multivariate .And Kaplan‐Meier method analysis of survival in the non‐small cell lung cancer patients with bone me‐tastases .Results Out of 383 patients with bone metastases ,178 patients with SREs .The incidence was 46 .5% .Univariate analysis showed that women ,adenocarcinoma ,never smoking history ,single bone metastases ,bisphosphonate therapy ,targeted therapy in patients with bone metastases are less likely to have SREs ,it was considered statistically significant (P<0 .05) .Multivariate analy‐sis showed multiple bone metastases and no bisphosphonate therapy is independent risk factors for SREs .Median survival time was 14 .5 months in non‐small cell lung cancer patients with bone metastases ,1 year survival rate was 46 .5% ,2 years survival rate was 15 .9% .The survival analysis shows that more bisphosphonate treatment and bisphosphonate with EGFR‐TKI therapy on the prog‐nosis of patients with statistically significance (P<0 .05) .Conclusion It was likely to occur SREs in NSCLC patients with bone metastases .No bisphosphonate and multiple bone metastases are independent risk factors for SREs .Bisphosphonate treatment may prevent or reduce occur SREs for NSCLC patients with bone metastases ,and it may prolong survival ,it speculated that bisphospho‐nate may have resistant NSCLC cell activity .

OBJECTIVETo investigate the roles of nitric oxide (NO) and eNOS in the pathogenesis of vasovagal syncope (VVS).

METHODSFourteen children with VVS (group A), 10 children with syncope other than vasovagal (group B) and 20 healthy volunteers (group C) were enrolled. Plasma NO levels in groups A and B were determined before and at the termination of the head-up tilt table test (HUT). The G894T polymorphism within the eNOS gene was determined in the three groups.

RESULTSPlasma NO levels in group A increased significantly when syncope attacked from 76.7+/-9.6 micromol/L (before HUT) to 90.0+/-11.4 micromol/L (P<0.05). After the syncope attack was improved, plasma NO level in group A was significantly reduced. There were no statistical differences in plasma NO levels before and after the HUT in group B. Determining the G894T polymorphism within the eNOS gene showed that group A was associated with a higher incidence of the GT gene type as compared to groups B and C (42.9% vs 10%; P<0.05).

CONCLUSIONSPlasma NO may be involved in the pathogenesis of VVS. The increased plasma NO level may be associated with the G894T polymorphism of the eNOS gene.

Child ; Humans ; Nitric Oxide ; blood ; physiology ; Nitric Oxide Synthase Type III ; genetics ; physiology ; Polymorphism, Genetic ; Syncope, Vasovagal ; etiology

ABSTRACT:Objective To investigate the clinical features of seronegative rheumatoid arthritis (RA)in western China and its outcomes after one-year treatment with disease modifying anti-rheumatoid drugs (DMARDs) so as to provide evidence for effective therapy.Methods We made a retrospective analysis of 240 RA patients treated in our department from May 2013 to June 2014.We compared the 47 seropositive and 25 seronegative RA patients in clinical features,laboratory parameters and outcomes after one-year DMARDs medication.Results The percentage of seronegative RA was 10.4% (25/240).The number of swollen small joints was significantly smaller in seronegative RA group (P<0 .0 1 ).Compared with those in seropositive RA,the level of hemoglobin was lower,the level of platelets was higher,and the level of alkaline phosphatase was lower in seronegative RA (P<0 .0 5 ).The remission rate was higher in seronegative RA group than in seropositive RA group after one-year DMARDs administration (P<0.05).Conclusion Seronegative RA is not rare in clinic.Even though seronegative RA patients often present fewer swollen small joints, it is difficult to distinguish between seronegative and seropositve RA just based on the clinical features.Besides,hematological damage is more severe in some patients with seronegative RA.Only after one-year treatment with DMARDs,the remission rate is higher in seronegative RA patients than in seropositve RA ones.

Objective To investigate the clinical characteristics of patients with polymyositis(PM) and dermatomyositis (DM), and compare the differences of PM/DM to help the understanding of clinical diagnosis and treatment. Methods One hundred and forty-five hospitalized PM/DM patients from Department of Rheumatology of the First Affiliated Hospital of Xiˊan Jiaotong University were collected from May 2008 to December 2014, and the clinical manifestations, muscle enzymes, electromyogram, muscle biopsy, treatment outcome were retrospectively analyzed. Mann-Whitney U test and χ2 test were used for statistical analysis. Results The most common initial symptom of PM was muscle weakness, accounted for 51.2%, while rash was the initial presentation in most DM patients(43.1%). The incidence of interstitial lung disease (ILD) (62.7% vs 39.5%, χ2=11.009, P=0.001), and the elevation of CRP (48.9% vs 26.8%, χ2=10.272, P=0.001) were all higher in DM than PM, while the elevation of level of CK (85.4% vs 61.8%, U=-2.668, P=0.008) and CKMB (82.9%vs 41.2%, U=-3.303, P=0.001) were more common in PM compared with DM. The pathological study showed degeneration of muscle fiber, connective tissue hyperplasia in most PM patients, and perimysium atrophy, vacuoles degeneration, muscle bundles, perivascular inflammatory cell infiltration were observed in most DM patients. During the follow-up, the clinical remission rate was 57.5%, the relapse rate and the mortality rate was 7.5%and 31.1%respectively. The mortality rate was higher in DM than PM (34.6% vs 21.4%, χ2=4.861, P=0.027). Infection and tumors were the major causes of death, and the lung was the most common site of infection. Conclusion Differences in the clinical features, muscle enzymes, CRP level, pathology and the mortality rate between PM and DM are evident, while ILD, infection and the higher mortality rate are more common in DM than in PM.

Chondroitinase has been used as an important tool in the study of the structure, function and distribution of glycosaminoglycans for many years. Recently, the enzyme has been reported to be a potential enzyme for chemonucleolysis, an established treatment for intervertebral disc protrasion. In this paper, a chondroitinase had been purified from the culture supernatant of Aeromonas sobria YH311 using a simple purification procedure of ammonium sulfate precipitation, QAE-Sephadex A50 ion exchange chromatography and Sephadex G-150 gel filtration. The immobilization of purified chondroitinase using sodium alginate or cellulose as carriers has also been studied. The chondroitinase obtained from Aeromonas sobria YH311 was purified 55-fold to 95.3% pure, the specific activity of the purified enzyme was 31.86u/mg and the yield was 37%. The molecular weight of chondroitinase from Aeromonas sobria YH311 was determined by SDS-PAGE to be 80kD, which was almost the same as those chondroitinase AC from Arthrobacter aurescens, Aeromonas liquefaciens and Flavobacterium heparinum. But its isoelectric point was 4.3 - 4.6, which was far lower than the microbial chondroitinase AC. After the immobilization on sodium alginate or cellulose, the properties of chondroitinase changed greatly. The optimum temperature and pH of the free enzyme were 50 degrees C and 7.0 respectively, and about 10% activity remained after heat treatment at 80 degrees C for 20 minutes, and 47% activity remained after two weeks storage at 4 degrees C. The chondroitinase immobilized on sodium alginate had the optimum temperature and pH of 40 degrees C and 7.0 respectively, about 50% activity remained after 80 degrees C heat treatment for 120 minutes and 50% remained after 30 days storage at 4 degrees C. The chondroitinase immobilized on cellulose had the optimum temperature and pH of 70 degrees C and 6.0 respectively, and more than 70% activity remained after heat treatment at 80 degrees C and 30 days storage at 4 degrees C. The yield of the immobilization was very low, with 18.56% for alginate and 18.86% for cellulose.
Aeromonas ; enzymology ; Chondroitinases and Chondroitin Lyases ; isolation & purification ; metabolism ; Enzyme Stability ; Enzymes, Immobilized ; metabolism ; Temperature

Objective To retrospectively determine the features of stones and calcifications in hepatobiliary system on virtual nonenhanced (VNE) dual-energy computed tomography (CT), and to evaluate the possibility of VNE images in diagnosis for those lesions.Methods A total of 128 gall stones and calcifications of the liver found in 110 patients were examined with triple phase abdominal CT scan from July 2007 to December 2011, in which true nonenhanced (TNE) phase and arterial phase were performed with single-energy CT (120 kVp) and portal venous phase was performed with dual-energy CT (100 kVp and 140 kVp). VNE images were generated from the portal venous phase dual-energy CT data sets by using commercially VNC software. The mean CT values for the stone, liver, bile and paraspinal muscle, mean lesion density and size in area dimension, contrast-to-noise ratio (CNR) of lesion to the liver or bile, and image noise were assessed and compared between VNE and TNE images. The effective dose and size-specific dose estimate (SSDE) were also calculated.Results The mean CT values of the lesions measured on VNE images declined significantly compared with those measured on TNE images (164.51±102.13 vs. 290.72±197.80 HU, P<0.001), so did the lesion-to-liver CNR (10.80±11.82 vs.18.81±17.06, P<0.001) and the lesion-to-bile CNR (17.24±14.41 vs. 21.32±17.31, P<0.001). There was no significant difference in size of lesions area between VNE and TNE images (0.69±0.88 vs. 0.72±0.85 cm, P=0.062). Compared to the 128 lesions found in TNE images, VNE images showed the same density in 30 (23.4%) lesions, lighter density in 88 (68.8%) lesions, while failed to show 10 (7.8%) lesions, and showed the same size in 61 (47.7%) lesions and smaller size in 57 (44.5%) lesions. The CT cutoff values of lesion and size were 229.21 HU and 0.15 cm, respectively. The total effective dose for triple phase scan protocol with TNE images was 19.51±7.03 mSv, and the SSDE was 39.84±11.10 mGy. The effective dose for dual phase scan protocol with VNE images instead of TNE images was 13.29±4.89 mSv, and the SSDE was 27.83±9.99 mGy. Compared with TNE images, the effective dose and SSDE of VNE images were down by 32.05%±3.69 % and 30.63%±2.34 %, respectively.Conclusions Although the CT values and CNR of the lesions decreased in VNE images, the lesions of which attenuation greater than 229.21 HU and size larger than 0.15 cmcould be detected with good reliability and obvious dose reduction. There was good consistency in the size of stones and calcifications in hepatobiliary system between VNE images and TNE images, which ensured the possibility of the clinical application of VNE images.

BACKGROUND:Abnormal activation of lymphocytes and nuclear factorκB-dependent non-specific inflammation are two major manifestations of joint damage in rheumatoid arthritis. Co-stimulatory signal CD40/CD40L is the dominant co-stimulatory factor in the recognition and activation of T cel s. IκBαeffectively inhibits nuclear factorκB pathway, prevent the inflammation in the central link, and suppress the damage caused by inflammatory factor in the synovial tissue.
OBJECTIVE:To investigate the therapeutic effect of double gene co-expressing adenovirus vector on arthritis based on an arthritis model rat transfected by CD40LIg-IRES2-IκBαco-expressing adenovirus vector.
METHODS:The pAdCD40LIg-IRES2-IκBαco-expressing adenovirus vector was established. Arthritic model was established through multi-subcutaneous injections of complete Freund's adjuvant of type col agen II (1 g/L) into Wistar rats. Then 20 arthritic rats were divided into two groups:untreated group and transfection group, receiving an injection of saline and pAdCD40LIg-IRES2-IκBαadenovirus vector to distal joint cavity of limbs, respectively.
RESULTS AND CONCLUSION:At 14 days post-transfection, compared with the untreated group, the mean arthritis index score, the CD40L expression of lymphocytes in synovial fluid, the nuclear factor-κB p65 expression in synovial tissue, and levels of interleukin-2, interleukin-6, tumor necrosis factor-α, matrix metal oproteinase-3 and matrix metal oproteinase-9 in synovial fluid of rats in transfection group were significantly lower than those in untreated group. Focal transfection of the CD40LIg-IκBαco-expression adenovirus vector can effectively inhibit arthritic symptoms, and reduce the expressions of inflammatory cytokine in synovial fluid and inflammatory molecule in synovial tissue of arthritic rats, which shows good therapeutic effect.

Objective CD4 +IL-17 +cells are a group of newly discovered effector CD4 +T cells, which may play a key role in the pathogenesis of cancer.This study aims to investigate the expres-sion of CD4 +IL-17 +cells in pancreatic cancer and its correlation with the clinicopathological characteristics and prognosis of the dis-ease as well as the clinical significance of the cells in the microenvironment of pancreatic cancer. Methods We collected tumor tis-sue and tumor-adjacent normal tissue samples from 51 pancreatic cancer patients.We determined the expressions of CD34 and vascular endothelial growth factor ( VEGF) and measured the proportion of IL-17 +cells in the cancer tissue using immunohistochemistry and the fluorescence activated cell sorter, respectively, followed by analysis of their correlation with tumor angiogenesis, clinicopathological pa-rameters, and survival time of the patients. Results The percentage of CD4 +IL-17 +cells in tumor tissue was positively correlated with microvessel density (r =0.534, P<0.05) and the expression of VEGF in the tumor tissue (r=0.356, P<0.05).IL-17 +cells were expressed more highly in the tumorous than in the tumor-adjacent normal tissue (P<0.05), and the expression level was correla-ted with the stage of tumor, node, and metastasis (TNM) and lymph node metastasis (P<0.05), but not with the patients′gender or age, tumor size, tumor location, histological grade, or local invasion (P>0.05).Fifty (98.0%) of the patients were successfully followed up for 2-67 months, which revealed a median survival time of 16.6 ±4.8 months, significantly longer in those with a higher expression of intratumoral IL-17 +cells (P<0.05).Univariate analysis showed an association of the survival rate with the tumor size, TNM stage, lymph node metastasis, and level of intratumoral IL-17 +cells, while multivariate analysis showed the TNM stage to be an independent prognostic factor for the survival of the pancreatic cancer patients. Conclusion The expression of CD4 +IL-17 +cells in the tumor tissue is positively correlated with tumor angiogenesis, while that of IL-17 +cells with the clinicopathological parameters and survival time of the patients and therefore may serve as an important immune indicator for the prognosis of pancreatic cancer.