Objective To explore the role of serum neutrophil extracellular traps (NETs) in rheumatoid arthritis (RA).Methods The serum circulating free DNA (cf-DNA) / NETs in 45 RA patients and 18 healthy controls were detected by Pico Green dsDNA Quantitation Kits,and the association between serum NETs and ESR,DAS28 score,anti-CCP antibodies was analyzed.T test,rank sum test,Pearson or Spearman correlation analysis were ued for statistical analysis.Results ① The levels of serum dsDNA/NETs in RA patients [0.523 0(0.282 0,1.637 0) μg/ml] were significantly higher than those in healthy controls [0.410 5 (0.140 0,0.966 0)μg/ml] (Z=-2.419,P=0.016).② The level of serum dsDNA/NETs in RA was positively correlated with ESR and DAS28 score (r=0.357,P=0.016; r=0.325,P=0.029),but not with anti-CCP antibodies(r=0.146,P=0.434).③ The levels of serum dsDNA/NETs in RA treated by DMARDs were lower than those of untreated with DMARDs[0.516 5(0.282 0,1.637 0) μg/ml,0.523 0 (0.369 0,1.485 0) μg/ml,Z=-1.215,P=0.225],but the difference was not significant.Conclusion NETs maybe associated with disease activity and play an important role in RA pathogenesis.

Objective To increase the understanding in protein-losing enteropathy (PLE).Methods Sixty-one PLE patients were enrolled in the study and the clinical characteristics, complicated disease, diagnosis and treatment were analyzed. Results The age of the patients was 16-77 (40±15)years, and the gender ratio was 35:26 (female: male). The main clinical manifestations were bilateral lower limb edema in 51 cases, ascites in 41 cases, bilateral pleural effusion in 23 cases, pericardial effusion in 13cases, abdominal pain in 16 cases and diarrhea in 33 cases. The prominent abnormality in laboratory examinations was hypoalbuminemia. The underlying diseases include systemic lupus erythematosus (SLE) in 28 cases, intestinal lymphangiectasia in 12 cases, hepatic cirrhosis in 5 cases, heart diseases in 5 cases,Crohn's disease in 3 cases, membranous nephropathy in 2 cases, Budd-Chiari syndrome in 1 case. Four cases happened after abdominal operation and 1 case after radiation therapy of gastric cardia cancer. Thirtyseven cases were diagnosed by 99Tcm-labelled human serum albumin scintigraphy and 24 cases were diagnosed clinically. Treatment was focused on underlying diseases. The clinical manifestations in 21 cases of SLE improved after SLE was controlled. In 2 cases of intestinal lymphangiectasia and one with Crohn's disease, the clinical manifestations improved after surgery. The other patients had no improvement.Conclusions PLE was not uncommon in clinical practice. Its predominant characteristics were severe hypoalbuminemia, edema and dropsy of serous cavity. PLE can complicate other diseases such as SLE,intestinal lymphangiectasia. Treatment should be focused on primary disease.

To reveal the underlying mechanism of doxorubicin induced hepatotoxicity, an NMR-based metabolomic approach combined with multivariate statistical analysis was used to observe its metabolic alternations of rat liver. Sixteen differential metabolites between model rats and normal rats were characterized as potential pathological biomarkers related to doxorubicin induced hepatotoxicity. Six pathways, including phenylalanine, tyrosine and tryptophan biosynthesis, valine, leucine and isoleucine biosynthesis, phenylalanine metabolism, glycine, serine and threonine metabolism, alanine, aspartate and glutamate metabolism, and tyrosine metabolism were regarded as the targeted metabolic pathways according to Metabolic Pathway Analysis (MetPA). The results suggested that the metabolic perturbations in rats with doxorubicin induced hepatotoxicity were mainly involved in amino acid metabolism, lipid pathways, purine metabolism, energy metabolism, dysfunction of biotransformation and oxidative stress. The investigation revealed the effects of doxorubicin on liver in a holistic metabolic way, which laid a foundation for further studies on its toxicity mechanism.
Animals ; Biomarkers ; metabolism ; Doxorubicin ; toxicity ; Energy Metabolism ; Liver ; drug effects ; metabolism ; Magnetic Resonance Imaging ; Magnetic Resonance Spectroscopy ; Metabolic Networks and Pathways ; Metabolomics ; Multivariate Analysis ; Oxidative Stress ; Rats

Objective To explore the significance of aryl hydrocarbon receptor (AhR) in patients with rheumatoid arthritis (RA) through detecting the levels of AhR and its response gene cytochrome P4501 A1 (CYP1 A1) in peripheral blood mononuclear cells (PBMCs).Methods Peripheral blood samples were collected from 35 patients with RA and 20 healthy subjects.The expression of AhR and CYP1A1 at mRNA level were detected by real-time PCR.The percentages of AhR-positive cells in PBMCs were detected by flow cytometry (FCM).The effects of leflunomide (LEF) on the expression of AhR and CYP1A1 were analyzed.The detailed clinical data of RA patients were recorded.The disease activity score (DAS) was calculated.Correlation analysis between AhR/CYP1A1 level and clinical data was conducted.Results (1) Both the expression of AhR at mRNA level and the percentage of AhR-positive cells in PBMCs from RA patients without LEF treatment were significantly higher than those from healthy subjects [(3.61±1.65) vs.(2.00±1.27),P=0.002; (34.21±11.30)％ vs.(18.83±7.32)％,P＜0.01].There were no statistically significant differences in the expression of AhR at mRNA level and the percentages of AhR-positive cells between patients with or without LEF treatment [(3.83 ± 1.62) vs.(3.61 ± 1.65),P =0.670 ; (36.69±10.61)％ vs.(34.21±11.30)％,P=0.462].(2) Non-LEF treatment group showed a higher relative expression of CYP1 A1 at mRNA level than that from control group [1.33 (0.08,7.86) vs.(0.62 ±0.29),z=-3.922,P＜0.01],but there was no statistical difference between LEF treatment group and non-LEF treatment group [(2.62±2.08) vs.1.33(0.08,7.86) z=-0.133,P=0.894].(3) Neither the expression of AhR and CYP1A1 at mRNA level nor the percentages of AhR-positive cells showed significant correlations with clinical data.Conclusion AhR was highly expressed in PBMCs from patients with RA,which might participate in the progression of rheumatoid arthritis.But the high expression of AhR did not reflect disease activity.Moreover,the treatment of LEF showed no significant influences on the expression of AhR and CYP1A1 in PBMCs from patients with RA.

Objective To analyze the failure ratio and the causes of the inoculation failure of hepatitis B virus(HBV)-vaccine in children and relevant the remedial measures. Methods One thousand three hundred and sixty cases treated in Suzhou Wuzhong people′s hospital during Jan.2007 to Jul.2008 were chosen,of whom 286 children from 1-5 years old to be anti-HBs negative or anti-HBs titre to be 0-10 IU/L were screened,and specific failure reasons for the vaccination were analyzed,also the timely treatment measures were taken.Then 286 children were divided into 5 groups randomly.Apart from one group was set up as blank control,the other 4 groups were arranged to accept different immunization methods with 0,1,2 month schedule,group A simply got revaccinated with HB vaccine(10 ?g) 3 times;group B revaccinated with double dosage of HB vaccine(20 ?g) 3 times;group C besides being revaccinated 3 times,the immune regulatory agent was jointly used;group D revaccinated 3 times with genetically engineered CHO hepatitis B vaccine. Results The ratio of failure of HBV-vaccine was 21.03%,what caused failure of hepatitis B vaccine included immunologic inadequacy 218(76.22%),repeated respiratory infection 192 cases(67.13%),abuse hormone 140 cases(48.95%),zinc deficiency 129 cases(45.10%),anaemia 108 cases(37.76%),passive smoking 80 cases(27.97%),the mother being chronic parenchymatous nephritis or HBV carrier 63 cases(22.03%),premature 54 cases(18.88%),adiposity 38 cases(13.29%),dystrophy 29 cases(10.14%).There were 4 methods of revaccination,the positive rate for group A,B,C,D were 90.00%,96.47%,99.08%,95.83%,respectively.Group C had the highest positive rate,compared with the other 3 groups,which were statistically significant(P a

Objective To evaluate magnifying endoscopy combined with narrow-band imaging ( ME-NBI) for diagnosis of early gastric cancer (EGC).Methods A total of 150 focal lesions from 143 patients over 35 years old identified by white light endoscopy (WLE) from March 2010 to December 2010 in our tertiary referential institution were recruited in the prospective study with written informed consent.Focal lesions were defined as any small local mucosa with abnormal shape or color based on an assessment of findings of WLE without any specified criteria, including superficial, depressed and elevated lesions.The patients with local advanced gastric cancer, submucosal lesions and history of gastrectomy were excluded from the study.All the patients received ME-NBI.Based on literature, national criteria of early diagnosis with ME-NBI were established.All the lesions underwent biopsy and pathological examination.Diagnostic accuracy of ME-NBI for EGC was assessed with reference to histopathology.Results In 150 focal lesions, 19 were pathologically diagnosed as EGC, 8 of which were treated by endoscopic resection and 11 were resected surgically.The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and accuracy of conventional WLE for diagnosing EGC were 94.7%, 53.4%, 22.8%, 98.6% and 58.7%, respectively.The counterparts of ME-NBI for diagnosing EGC were 73.7%, 99.2%, 93.3%, 96.3% and 96.0%, respectively.The diagnostic accuracy of ME-NBI was significantly better than that of conventional WLE (96.0% vs.58.7%, P＜0.05).With regard to the findings of EGC on ME-NBI, irregular or absent microsurface pattern and microvascular pattern were characteristic features of EGC.Conclusion Conventional WLE is still an important and mandatory screening modality, which is significant for further procedures of suspected lesions, preferably accompanied with biopsy.ME-NBI achieved superior accuracy in the differential diagnosis of focal lesions detected with conventional WLE, but needs further verification.

BACKGROUNDGingerol is the generic term for pungent constituents in ginger, which has been reported to be effective for inhibiting vomiting. We attempted to investigate the antiemetic effect of gingerol and its effective mechanism on substance P and NK(1) receptors in minks.

METHODSThe antiemetic effect of gingerol was investigated during a 6-hour observation on a vomiting model in minks induced by cisplatin, (7.5 mg/kg, intraperitoneal). The distribution of substance P and NK(1) receptors in the area postrema and ileum were measured by immunohistochemistry, and the expression of NK(1) receptor in the area postrema and ileum were measured by Western blotting.

RESULTSThe frequency of cisplatin induced retching and vomiting was significantly reduced by pretreatment with gingerol in a dose-dependent manner (P < 0.05). Substance P-immunoreactive was mainly situated in the mucosa and submucosa of the ileum as well as in the neurons of the area postrema. The immunoreactive production of NK(1) receptor was mainly situated in the muscular and submucosa of ileum and the neurons of area postrema, gingerol markedly suppressed the increased immunoreactivity of substance P and NK(1)1 receptor induced by cisplatin in a dose-dependent manner (P < 0.05), and exhibited effective inhibition on the increased expression levels of NK(1) receptor in both the ileum and area postrema dose-dependently (P < 0.05).

CONCLUSIONSGingerol has good activity against cisplatin-induced emesis in minks possibly by inhibiting central or peripheral increase of substance P and NK(1) receptors.

Animals ; Area Postrema ; metabolism ; Blotting, Western ; Catechols ; therapeutic use ; Disease Models, Animal ; Fatty Alcohols ; therapeutic use ; Ileum ; metabolism ; Immunohistochemistry ; Male ; Mink ; Receptors, Neurokinin-1 ; metabolism ; Substance P ; metabolism ; Vomiting ; chemically induced ; drug therapy

The aim of this study was to verify the presence of multipotential mesenchymal stem cells in peripheral blood (PBMSC) of rabbits. For mobilization, granulocyte-colony-stimulating factor 30 µg/(kg·d) was injected into New Zealand White rabbits subcutaneously for 6 d, then the PBMSC were isolated from peripheral blood of rabbits by density gradient centrifugation and adhesive culture. The morphology of cell proliferation was observed by microscopy, the proliferative curve of cells was drawn. The phenotypes of PBMSC were detected by flow cytometry, the differential capability of PBMSC into osteocytes, chondrocytes and adipocytes was identified. The results showed that the morphology of subcultured PBMSC were spindle or polygonal shaped, and cell population doubling time was 37.4 h. The isolated PBMSC expressed mesenchymal marker CD29, but not expressed hematopoietic marker CD14. Under specific induction conditions, PBMSC demonstrated multipotency to differentiate into osteocytes, chondrocytes and adipocytes. It is concluded that PBMSC are successfully isolated from peripheral blood and cultured, and their multipotential capability of differentiation into osteocytes, chondrocytes and adipocytes are verified.
Adipocytes ; cytology ; Adipogenesis ; Animals ; Animals, Newborn ; Cell Differentiation ; Chondrocytes ; cytology ; Mesenchymal Stromal Cells ; cytology ; Multipotent Stem Cells ; cytology ; Osteocytes ; cytology ; Rabbits

OBJECTIVETo investigate drug resistance status in patients with highly active antiretroviral therapy (HAART) in Shandong province.

METHODSA total of 758 patients were separated from the anticoagulatory whole blood during May and October in 2011. The entire protease gene and part of the reverse transcriptase gene were amplified by RT-PCR and nest-PCR in the samples with viral load larger than 1000 copies/ml, then sequenced the gene fragments. Mutation of drug resistant gene and drug susceptibility was analyzed by the online tool HIV db program developed by Stanford University.

RESULTSThe rate of virologic failure in patients was 9.1% (69/758). A total of 53 gene sequences that acquired were used for genotypic resistance analysis. A total of 23 patients were indicated drug resistance with the total of 3.1% (23/742). Drug resistance rates of nucleotide reverse transcriptase inhibitor (NRTI) and non-NRTI(NNRTI) were 2.4% (18/742) and 3.0% (22/742), respectively, and the primary mutation types of drug resistance were M184V and Y181C for NRTI and NNRTI, with no resistance to protease inhibitor (PI). In the 23 patients indicated drug resistance, 78.3% (18/23) were NRTI resistance, 95.7% (22/23) were NNRTI resistance and 73.9% (17/23) dual NRTI and NNRTI resistance.

CONCLUSIONThe presence of drug resistant gene in HIV strains among AIDS patients with HAART in Shandong province was at low level, but mutation diversity was found in drug resistant gene.

Acquired Immunodeficiency Syndrome ; drug therapy ; virology ; Adolescent ; Adult ; Aged ; Antiretroviral Therapy, Highly Active ; Drug Resistance, Viral ; genetics ; Female ; Genes, Viral ; Genotype ; HIV-1 ; drug effects ; genetics ; Humans ; Male ; Middle Aged ; Mutation ; Sequence Analysis ; Viral Load ; Young Adult

OBJECTIVEIn order to develop the diagnostic genechip for specific detection of Schistosoma japonicum (Chinese mainland strain).

METHODSProbe and primers were designed based on the Schistosoma japonicum 5D gene encoding an immunogenic miracidial antigen. The probe for the conservative and specific gene sequence was spotted onto the specially treated glass slides by pin-based spotting robot Pixsys 5500 and was employed to make genechips. A polymerase chain reaction (PCR) protocol was designed to effectively amplify the 5D gene fragment containing the probe sequence from cercaria, egg, adult worm and infected Oncomelania DNA as well as other flukes DNA, respectively. After 35 cycles by PCR, the products were then labeled with fluorescent Cy3-labeled primer, using dissymmetrical PCR. The labeled PCR products of the target genes were hybridized to the diagnostic genechips for detection of Schistosoma japonicum and a fluorescent scanner (ScanArray 3000) was used to observe and record the hybridization signals.

RESULTSThe result obtained from the study showed that a 262 bp DNA fragment was amplified from cercaria, egg and adult worm with the designed primers and enable the genechip be applied to detect a single cercaria, egg and adult worm. When the genechip was used to detect Clonorchis sinensis, Fasciolopsis busk, and Paragonimus westermani DNA, the results showed negative, indicating that the genechip had good specificity.

CONCLUSIONThe genchip technique for detection of Schistosoma japonicum was established successfully and having the characteristics of high sensitivity and specificity.

Animals ; China ; DNA, Helminth ; genetics ; Genes, Helminth ; genetics ; Genetic Techniques ; Polymerase Chain Reaction ; methods ; Schistosoma japonicum ; genetics ; Sensitivity and Specificity