AIM: To study the expression of nucleostemin (NS) gene in human breast tumor tissues and the relations of NS gene expression level with histological grades, histological types and TNM stages of the tumor.METHODS: Total RNA was isolated from human breast tumor tissue. The methods of electrophoresis and RT-PCR were used in measuring NS gene expression level, and the relations of NS gene expression level with histological grades, histological types and TNM stages of the tumor were analyzed. RESULTS: The results indicated that there was no NS gene expression detected in normal breast tissues, and NS gene expression in malignant breast tumor tissues (P0.05). CONCLUSION: It is suggested that there is no relation of NS gene expression level with histological types of the breast cancer, but there is a marked correlation of NS gene expression level with the histological grades and TNM stages.

Objective:To evaluate the clinical performance of low-count platelet(PLT) samples for Mindray BC-6800 automatic hematology analyzer and compare the detection results between two sampling mode(automatic injection, manual injection).Methods: To evaluate the repeatability and contamination rate of low-count platelet samples depend on ICHC, CLSI and other standards; to choose 108 blood samples with thrombocytopenia and detect them in two kinds of detection mode (automatic and manual operation), and then evaluate the consistency of low-count PLT samples between the two modes.Results: The results of two detection modes for low-count platelet samples showed there were well repeatability between them. The CV% of those samples which PLT were upon 20×109/L can be assurance within 5% in two modes. The value and fluctuation of SD and range of those samples which PLT were under 20×109/L were smaller; the SD of two modes were 0.5 and 0.8, respectively; and the Range were 1 and 2, respectively. Both of the two modes were less than 1% in carryover rates, high PLT accounts has no influence on low accounts. The correlation coefficient was up to 0.9910 between the two modes by analyzing the 108 blood samples. The results demonstrate there were low deviation and well consistency between the two modes by analyzing medical decision level concentration point for them.Conclusion: Mindray BC-6800 has good clinical performance to detect low-count PLT samples and there are well consistency between automatic and manual operation. Clinical Laboratory chooses which suitable mode to detect samples may depend on self-situation.

The national examination of doctors' qualification is the examination which assesses the medical students' knowledge and skills for doctors' work. This paper introduced autorities concerned, implementing agencies, examination forms and procedures, examination contents and keypoints, pass-ing criteria and matters needing attention of national examination of doctors's qualification be tween Canada and Australia expecting to provide references for the revolution of Chinese Examination of Doctors' Qualification.

Objective To explore the clinical and serological features of patients with pneumomdiastinum (PNM) and dermatomyositis associated interstitial lung disease (DM-ILD).Methods A total of 145 polymyositis/dermatomyositis (PM/DM) patients hospitalized in our department from March,2010 to December,2012 were recruited.The sera,clinical and laboratory data were collected.Anti-aminoacyl-tRNA synthetase (anti-ARS) and anti-melanoma differentiation-associated gene 5 (MDA5) antibodies were detected by RNA-immunoprecipitation (RNA-IP) and ELISA,respectively.Anti-NXP2,anti-TIF1γ,anti-SRP,anti-SAE and antiMi2 antibodies were detected by immunoprecipitation-Western blotting.Variables were compared between DM patients with and without PNM,as well as between DM-ILD patients with and without PNM.Chi square test,Fisher's exact test,t test,Mann-Whitney U test and a multivariate logistic regression model were used for statistical analysis.Results A total of 11 DM patients (10 patients positive for anti-MDA5 and 1 patient positive for anti-Mi2) developed spontaneous PNM.No PM patient developed PNM.No differences of sex,age at the onset of DM,serum ferritin levels and C reactive protein (CRP) levels was observed between DM patients with and without PNM.Compared with DM patients without PNM,DM patients with PNM had significantly higher frequencies of rapidly progressive ILD (RP-ILD) (63.6％ vs 24.4％,x2=7.25,P=0.01),anti-MDA5 antibodies (90.9％ vs 52.4％,x2=5.86,P=0.02),clinically amyopathic DM (CADM) (63.6％ vs 22.0％,x2=8.57,P=0.007) and cutaneous ulcers (36.4％ vs 11％,x2=5.20,P=0.04),but significantly lower creatine kinase (CK) levels [58.5 (30.5,394.3) U/L vs 284 (73.0,917.0) U/L,t=207.5,P=0.04].The logistic multivariate analysis indicated that cutaneous ulcer was the only independent risk factor for the occurrence of PNM in DM [OR=5.98,95％ confidence interval (CI) (1.12,31.98),P=0.037].In the comparisons between DM-ILD patients with and without PNM,a higher frequency of CADM was observed in the PNM group (63.6％ vs 27.9％,x2=5.37,P=0.03),but no significant difference of the frequencies of RP-ILD,anti-MDA5 antibodies,cutaneous ulcers and CK levels was found in these two groups.All patients with PNM were treated with corticosteroids and immunosuppressants.During the follow-up period,6 patients died of respiratory failure.Conclusion Spontaneous PNM is a refractory complication with poor prognosis,and tends to occur in DM patients with RP-ILD,anti-MDA5 antibody,CADM diagnosis and low CK level,especially in patients with cutaneous ulcers.

Objective:To discuss the autoimmune response on the mechanism and clinical significance of nerve root injury induced by the non-compressive nucleus pulposus protrusion.Methods:Forty-eight female SD rats were randomly divided.Twenty-four female SD rats were recruited as model group,and the others as control.After ten days,twenty days and forty days,the pain threshold of left hind leg and the levels of TNF-α, CD4+, CD8+T cells were measured, the nerve root changes in morphology were observed by HE.Results:Compared with the control group,model group in ten days,twenty days,which left hind leg pain threshold,the proportion of CD4,CD8+T cell and the expression of TNF-αare significantly different.After forty days,these two groups have no statistical differ-ence.After ten days,lumbar nerve root cross-sectional myelin of model group was partial disintegrated.The worst damage happened in twenty days,and almost recovered to normal in forty days.Conclusion:T cell-mediated autoimmunity and TNF-αplay an important role to the nerve root injury in the early time of the non-compressive nucleus pulposus protrusion.

BACKGROUND:Superparamagnetic iron oxide (SPIO) labeling can trace the migration of stem cells in vivo, and the fluorescent DiI dye is suitable for marking and tracing cells because of its less influence on cellviability, proliferation and differentiation.
OBJECTIVE:To observe the effect and safety of SPIO and fluorescent DiI dye to double label bone marrow mesenchymal stem cells.
METHODS:The bilateral lower limbs of rats were isolated sterilely. Bone marrow was obtained by rinsing using low-glucose DMEM. Bone marrow mesenchymal stem cells were isolated by the whole bone marrow adherence method and purified by differential attachment method. Purified cells were dual-labeled with SPIO particle and fluorescent DiI dye.
RESULTS AND CONCLUSION:Bone marrow mesenchymal stem cells could be separated at 8-10 days after primary culture and the subculturing cycle was 3-4 days. Rat bone marrow mesenchymal stem cells could be effectively labeled with SPIO-DiI and the labeling efficiency was almost 100%. Blue irons contained in intracytoplasmatic vesicles could be observed clearly with Prussian blue staining, and the fluorescence microscopy showed red fluorescence at cytoplasm. Survival and apoptosis percentages obtained by MTT analysis were similar among labeled and unlabeled bone marrow mesenchymal stem cells that were both about 95%.These findings indicate that the rat bone marrow mesenchymal stem cells could be efficiently labeled with SPIO-DiI to construct a nano-bioprobe, without significant changes in morphology, viability and proliferation.

Objective To investigate the expressions of FAT10 and p53 mutant in gastric cancer tissues and their relations. Methods Immunohistochemistry and RT-PCR were used to detect the expressions of FAT10 and p53 in gastric cancer tissues (n=62), para-cancerous tissues (2-5 cm apart from cancer, n=62), and normal gastric tissues (7>5 cm apart from cancer, n=62). The association of FAT10 with p53 and clinical outcomes were analyzed by Spearman and Pearson correlation. Results The immunohistochemistry examination showed that expressions of FAT10 [51.61%(32/62)] and p53 [45.16% (28/62)] were significantly higher in cancerous tissues than in para-cancerous tissues [12.90%(8/62) and 14.51% (9/62), χ2=21.26 and 20.69, P<0.01] and normal tissues [6.45% (4/62) and 9.68% (6/62), χ2=13.91 and 19.61, P<0.01]. Overexpressions of FAT10 protein and mRNA in cancerous tissues were closely related to lymph node metastasis and TNM staging (both P value<0.05). There was a positive correlation between FAT10 and p53 in protein and mRNA expressions (protein r=0. 865, P<0.05; mRNA r=0.761, P< 0.01). Those with positive expression of FAT10 had lower survival rate compared to those with negative expression (P<0.05). Conclusions The positive relation between over-expression of FAT10 and p53 implicates that both are involved in the gastric carcinogenesis, and FAT10 is a novel gastric cancer marker with prognostic significance.

Objective To investigate the clinical features of polymyositis/dermatomyositis (PM/DM) with anti-nuclear matrix protein (anti-2NXP2) antibodies in Han-Chinese.Methods ImmunoprecipitationWestern Blotting (IP-WB) method was used for screening anti-NXP2 antibodies in 141 adult Han-Chinese patients with PM/DM.The clinical and laboratory data were collected,analyzed and compared with the antiNXP2-positive patients reported in the literature.Statistical analyses were performed using chi-square test and Fisher's exact test.Results Seven (5％) patients,including 6 with DM and 1 with PM,were identified as anti-NXP2 positive.Consistent with the 114 anti-NXP2-positive patients reported in the literature,Han patients with anti-NXP2 had higher frequencies of weakness,heliotrope rash and Gottron's sign.Compared with anti-NXP2-negative patients,anti-NXP2-positive patients presented significantly higher frequencies of dysphagia [43％(3/7) vs 9％(12/134),x2=8.04,P=0.027] and edema [43％(3/7) vs 2％(3/134),x2=26.94,P=0.001 4],while an absence of PM/DM related cancer was observed in Han patients with anti-NXP2.Conclusion Adult Han-Chinese patients with anti-NXP2 are DM predominant,and are characterized by a high frequencies of edema and dysphagia.

Objective To investigate the effect of unfractionated heparin (UFH) on the expression of heme oxygenase-1 (HO-1) in intestinal mucosa of mice with sepsis.Methods Thirty-six male C57BL/6J mice were randomly divided into sham group,cecal ligation and puncture (CLP) group and UHF group,n =12 in each group.Model of intestinal injury in sepsis was induced by CLP.In sham group,the mice were exposed without ligation of cecum.In UFH group,the mice were treated intravenously with 8 U of UFH via the tail vein half an hour before the operation and 12 hours after the surgery respectively.Six mice in each group were randomly chosen at 4 hours and 24 hours after operation to collect inferior vena venous blood samples and terminalileum tissues.The serum levels of interleukins (IL-1 β,IL-6),and tumor necrosis factor-α (TNF-α) were determined by enzyme linked immunosorbent assay (ELISA).The serum level of D-lactate was determined by colorimetry.Pathological changes of ileum tissue and Chiu score were observed after hematoxylin eosin (HE) staining.The HO-1 expression was detected immunohistochemically.Results In sham group,no significant changes in the serum levels of IL-1 β,IL-6,TNF-α and D-lactate were observed.Twenty-four hours after the operation,the structure of intestinal mucosa was basically normal without obvious pathology change and no HO-1 positive cells were found.The serum levels of IL-1 β,IL-6,TNF-α,and D-lactate in CLP group were gradually increased,and they were significantly increased as compared with sham group [IL-1 β (ng/L):40.87±2.88 vs.22.60±2.05 at 4 hours,113.73±3.96 vs.22.07±2.74 at 24 hours;IL-6 (ng/L):63.89±3.26 vs.44.89±3.38 at 4 hours,176.56±5.45 vs.45.76±4.02 at 24 hours;TNF-α (ng/L):194.62± 14.13 vs.152.05±6.22 at 4 hours,599.62± 10.20 vs.155.90± 14.18 at 24 hours;D-lactate (mmol/L):0.24± 0.02 vs.0.19 ± 0.01 at 4 hours,0.33 ± 0.04 vs.0.20 ± 0.02 at 24 hours,all P ＜ 0.05].Twenty-four hours after the operation,edema and inflammation in ileal mucosa,intestinal villi structural damage were observed,the Chiu score was significantly higher than those in the sham group [4.5 (3.0-5.0) vs.0 (0-1.0),P ＜ 0.05],and a small amount of HO-1 positive cells were localized in the intestinal mucosa.Compared with CLP group,the serum levels of IL-1 β,IL-6,TNF-α,and D-lactate of UFH group were significantly decreased [IL-1 β (ng/L):31.53 ± 2.90 vs.40.87 ± 2.88 at 4 hours,61.13 ± 2.80 vs.113.73 ± 3.96 at 24 hours;IL-6 (ng/L):51.16 ± 5.68 vs.63.89 ± 3.26 at 4 hours,81.16 ± 4.54 vs.176.56 ± 5.45 at 24 hours;TNF-α (ng/L):171.76± 5.60 vs.194.62± 14.13 at 4 hours,328.48 ± 10.79 vs.599.62± 10.20 at 24 hours;D-lactate (mmol/L):0.21 ±0.01 vs.0.24±0.02 at 4 hours,0.24±0.02 vs.0.33±0.04 at 24 hours,all P ＜ 0.05].Twenty-four hours after the operation,intestinal injury was ameliorated,the Chiu score was significantly lower [1.5 (1.0-5.0) vs.4.5 (3.0-5.0),P ＜ 0.05],and HO-1 positive cells in the intestinal mucosa was remarkably increased.Conclusion UFH can enhance the expression of HO-1 in intestinal mucosa,reduce the release of inflammatory factors,ameliorate the intestinal inflammatory response,and thus play a protective role in intestinal tissue in mice with sepsis.