Objective:To detect anti-LKM-1 antibody with enzyme-linked immunosorbent assay (ELISA) using a recombinant fusion peptide which comprises 257-351 amino acid fragment of CYP2D6 as antigen.Methods:We obtained CYP2D6 cDNA fragment by means of PCR,using total liver cDNA library as the template.The PCR products were ligated into pEGH expressing vector to construct the recombinant expressing vector with high efficiency in Saccharomyces Cerevisiae Y258.The positive clones were identified by PCR reaction and then induced by galactose.Glutathione-Sepharose 4B was used for purification of recombinant CYP2D6 protein.After affinity purification,the antigenicity was identified with Western blot.Serum samples from 26 patients who were positive for anti-LKM-1 antibody,20 patients with other connective tissue disorder(CTD) and 30 normal controls were retrospectively tested with ELISA.Results:A fusion peptide was expressed and purified.The antigenicity was confirmed with Western blot using standard of anti-LKM-1 antibody-positive serum.Of the 26 serum samples which are positive for anti-LKM-1 antibody,5 of 6 samples positive for anti-HCV antibody also recognized the recombinant fusion peptide with ELISA,only one serum sample which was showed positive anti-HCV antibody displayed a negative result in ELISA assay.All other 20 patients with positiv anti-LKM-1 antibody were shown positive in ELISA assay using this recombinant peptide.All the serum samples from patients with other CTD were negative in ELISA assay.Conclusion:The recombinant antigen fragment contains major epitope regions in natural CYP2D6 antigen.Detection of anti-LKM-1 antibody with ELISA using the recombinant peptide can improve the sensitivity and has a potential role in determining its clinical association.

Objective To assess the levels of glucose-6-phosphate isomerase(GPI) mRNA in peripheral blood monocytes and serum GPI levels in patients with rheumatoid arthritis, and analyze the association of serum GPI with MCV antibody, CCP antibody and RF of RA. Methods Fluorogenic quantitative real-time polymerase chain reaction (FQ-RT-PCR) was used to examine mRNA expression on peripheral blood monocytes in 60 RA patients (28 case in active stage,32 cases in stable stage) ,30 patients with other rheumatic diseases, and 30 healthy controls. ELISA was used to detect the levels of serum GPI, anti-MCV antibodies, anti-CCP antibodies and RF in each group. Results The levels of GPI mRNA in RA group [△Ct=4.21 (3.04-7.23)] were significantly higher than those in patients with other rheumatic diseases [△Ct=8.42 (5.16-9.98),P<0.01] and healthy controls [△Ct=8.66 (4.90-10.01), P<0.01]. There were statistically significant differences of GPI mRNA levels between active RA [△Ct=3.78 (1.28-6.09)] and inactive RA[△Ct =5.88(3.23-8.94),H=11.760,P<0.01)]. The RA group serum GPI levels [3.02 (2.02-8.39) mg/L] were higher than those of other rheumatic diseases [0.20 (0.11-0.32) mg/L] and healthy controls [0.18(0.08-0.30) mg/L]. There were significant differences of serum GPI levels between active RA group [4.84(2.81-10.38) mg/L] and inactive RA group[2.12 (1.26-4.34) mg/L] (H=9.830, P<0.01). The sensitivities of GPI, anti-MCV and anti-CCP were 68% (41/60) ,57% (34/60),58% (35/60), respectively and specificities were 95% (57/60), 92% (55/60) and 93% (56/60), respectively. Conclusions The high expression of GPI mRNA in RA patients shows that it may play a pathological role in the development of RA, and it may be correlated with the activity of RA. It may be a valuable diagnostic parameter for RA, because of its high sensitivity and specificity.

Objective To investigate the diagnostic value of apparent diffusion coefficient in the evaluation of sinonasal masses.Methods Sixty-seven sinonasal solid masses over 1 cm in diameter confirmed by pathology were retrospectively analyzed,all patients underwent preoperative routine MRI with DWI,the ADC values were measured in ROI within the solid mass.The patients were divided into benign and malignant groups by the histopathology,according to pathological findings,the patients were further divided into the hematolymphoid tumors,the malignancy of epithelium and mesenchymal tissue,the benign tumors of epithelial and mesenchymal tissue,and vasogenic masses.ANOVA test and t test were used to compare the ADC values of different groups.The receiver operating characteristic curve (ROC) was constructed using various cut points of ADC for different parameters to confirm the diagnostic threshold value and evaluate the diagnostic efficacy.Results All lesions were solitary.There were 22 malignant tumors,of which 6 lesions were hematolymphoid tumors and 16 lesions malignancy from epithelium and mesenchymal tissue.There were 45 benign tumors,of which 22 lesions were benign tumors from epithelium and mesenchymal tissue and 23 lesions vasogenic masses.The mean ADC value of malignant and benign masses was(0.88 ± 0.26) × 10-3 mm2/s and (1.54 ± 0.41) × 10-3 mm2/s respectively.There was statistically significant differences between them (t =6.897,P ＜ 0.01).The mean ADC value was(0.63 ± 0.10) × 10-3 mm2/s in hematolymphoid tumors,(0.97 ±0.24) × 10 3 mm2/s in malignancy from epithelium and mesenchymal tissue,(1.38 ± 0.23) × 10-3 mm2/s in benign tumors from epithelium and mesenchymal tissue,(1.68 ± 0.49) × 10-3 mm2/s in vasogenic masses respectively.There was statistically significant difference among all 4 groups(F =22.788,P ＜ 0.01),and the differences between any 2 groups were still statistically significant(P ＜ 0.05).The area under the ROC calculated was 0.945.Using an ADC value of 1.08 × 10-3 mm2/s as the threshold value for differentiating malignant from benign lesions,the best result obtained had a sensitivity of 81.8％ (18/22),specificity of 97.8％ (44/45),accuracy of 92.5 ％ (62/67).Conclusion The ADC value is a valuable tool in differentiating benign from malignant masses and different kinds of masses in sinus and nasal cavity.

Objective To determine the CT and MRI features of the sinonasal cavity. Methods The CT and MRI of nine patients with histologically proved hemangiopericytoma in the sinonasal cavity were retrospectively reviewed. All nine patients underwent CT and seven of them underwent MRI. In addition, the time-intensity curve (TIC) of dynamic contrast-enhanced (DCE) MRI were analyzed in three patients. Results The lesions were from the nasal cavity in 5 cases, the maxillary sinus in 3 cases, and the sphenoid sinus in one case. The lesions appeared spindle in 4 cases, oval in 3, and irregular in 2. The mean maximum diameter of the lesions was 31 mm (range, 15 to 52 mm). Seven hemangiopericytomas had welldefined margins and 2 had ill-defined margins. On plain CT, the lesions were isodense to gray matter in six (66. 7% ) and slightly hyperdense in three (33. 3% ) patients. Only two patients had post-contrast CT and the tumors showed marked enhancement. The lesions caused adjacent bony compression and absorption in 7 cases and bony destruction in 2. On MR T1WI, hemangiopericytomas appeared hypointense in 3 (42. 8% ) patients and isointense in 4 (57. 2% ) patients. On T2WI, the lesions appeared hyperintense in three (42.8%) patients and isointense in four (57.2%) patients. On T1WI, all the lesions showed relatively homogeneous signal, while 2 lesions revealed heterogeneous signal on T2WI. Hemangiopericytomas showed marked homogeneous contrast enhancement in 4 cases and heterogeneous contrast enhancement in 3.TIC showed a steady enhancement pattern in 3 patients. The lesions involved the orbit in 2 cases and,cavernous sinus, anterior skull base meninge, pterygopalatine fossa and infratemporal fossa in one case. Five patients underwent follow-up for two to eight years after surgery. Two patients were found to have recurrence.Conclusions Marked enhancement and steady enhancement pattern of TIC on MRI are typical manifestations of hemangiopericytoma of the sinonasal cavity. Bone changes may help to identify the nature of the lesion. Imaging can accurately detect the scope of hemangiopericytoma in this region.

Objective To explore the features of inflammatory pseudotumor in the maxillary sinus on CT and MRI.Methods The CT and MRI data of 8 patients with histology-proven inflammatory pseudotumor in the maxillary sinus were retrospectively analyzed.All 8 patients underwent CT scans and 7 of them also underwent MRI examinations.In addition, the time-intensity curve (TIC) of dynamic contrast-enhanced (DCE)MR imaging were analyzed in 3 patients.Results Two cases had lesions in the left maxillary sinus, while the others showed lesions in the right maxillary sinus.All the lesions showed irregular shape with well-defined margins in 5 cases and hazy margins in 3 cases.The mean maximum diameter of the lesions was 51 mm (ranged from 29 mm to72 mm).On non-enhanced CT, the lesions were isodense to gray matter in 6 and slightly hypodense in 1 patients.Only 2 patients had post-contrast CT with moderate enhancement of their lesions.The lesions caused adjacent bony absorption and destruction with residual bony sclerosis.On MR T_1 WI, inflammatory pseudotumor showed hypointense in 2 patients and isointense in 5 cases in relative to gray matter.On T_2 WI, the lesions revealed inhomogeneous hypointense in 6 patients and isointense in 1 patients.Inflammatory pseudotumor showed heterogeneously slight contrast enhancement in 2 cases and moderate enhancement in 5 cases.The TIC showed a steady enhancement pattern in 3 patients.The lesions extended to nasal cavity in 6 cases, pterygopalatine fossa and infratemporal fossa in 4, orbit in 4, and cheek in 3 cases.Six patients underwent follow-up for 2-5 years after surgery and 2 of them were found to have recurrence.In comparison to CT, MRI was proved to show the extent of the lesions more clearly.Conclusions Bony destruction with sclerosis and hypointense signal on MR T_2 WI are typical manifestations of inflammatory pseudotumor in the maxillary sinus.Combined CT and MRI can provide clinicians with more comprehensive information for the diagnosis, therapy, and follow-up of these lesions.

Objective To study the protection of vanillin derivative VND3207 on the cytogenetic damage of mouse bone marrow cell induced by ionizing radiation.Methods BALB/c mice were randomly divided into five groups:normal control group,2 Gy dose irradiation group,and three groups of 2 Gy irradiaiton with VND3207 protection at doses of 10,50 and 100 mg/kg,respectively.VND3207 was given by intragastric administration once a day for five days.Two hours after the last drug administration,the mice were irradiated with 2 Gy γ-rays.The changes of polychromatophilic erythroblasts micronuclei (MN),chromosome aberration (CA) and mitosis index (MI) of mouse bone marrow cells were observed at 24 and 48 h after irradiation.Results Under the protection of VND3207 at the dosages 10,50,100 mg/kg,the yields of poly-chromatophilic erythroblasts MN and CA of bone marrow cells were significantly decreased(t = 2.36-4.26,P ＜ 0.05),and the marrow cells MI remained much higher level compared with the irradiated mice without drug protection (t = 2.58,2.01,P ＜ 0.05).The radiological protection effect was drug dose-dependent,and the administration of VND3207 at the dosage of 100 mg/kg resulted in reduction by 50% and 65% in the yields of MN and CA,respectively.Conclusions VND3207 had a good protection effect of on γ-ray induced cytogentic damage of mouse bone marrow cells.

Objective To study the CT and MRI findings of hemorrhagic and necrotic nasal polyps. Methods The imaging data of 17 cases with hemorrhagic and necrotic nasal polyps confirmed by surgery and pathology were analyzed retrospectively. CT was performed in 14 cases, MRI in 16 cases, of which 15 also underwent contrast-enhanced MRI. Results All 17 lesions with well-circumscribed margin originated in the areas of maxillary sinus ostia and extended into adjacent nasal cavity and maxillary sinus. The lesions appeared as lobular shape in 16 cases and oval shape in 1 case. On non-enhanced CT, 14 lesions showed heterogenous soft tissue density masses, the hyperdense strip and nodule were detected on the periphery and in the center of 2 lesions respectively. All the involved bony walls were compressed and remodeled with focal defect,especially in the medial wall of maxillary sinus. On MRI, all 16 lesions revealed inhomogenous signal. The central region of the lesions appeared hypointense signal on T_1WI compared to gray matter and hyperintense signal on T_2WI with line-like hypointense signal septa in 14 cases, the irregular hypointense signal rims were also found on the periphery of 15 lesions on T_2WI, postcontrast MR imaging showed strongly inhomogenous enhancing masses with non-enhancing hypointense rim, the appearances of enhancement showed multiple nodules in 10 cases, patches in 4 cases and leaf-like in 1 case. The typical simple polyps were present in the ipsilateral nasal cavity of the lesions in 4 cases, extending into nasal vestibule anteriorly and choana posteriorly. On dynamic contrast-enhanced MRI in 11 cases, the timeintensity curves (TIC) showed increasing enhancement type in 7 cases and rapid enhancement and slow wash-out type in 4 cases. Conclusion The inhomogenous hyperintensity surrounded by the peripheral hypointense rim on MR T_2WI and marked nodular and patchy enhancement appearance are typical features of hemorrhagic and necrotic nasal polyps. CT is helpful to judge the nature of lesions, but difficultly confirms the diagnosis, while MRI should be the imaging modality of first choice for hemorrhagic and necrotic nasal polyps.

Objective To investigate the growth and exocrine function of BM-MSC derived from PBC patients.Methods To compare the growth patterns and cytokines secretions between PBC patients and healthy controls by student's t test.Results ① There was no difference in growth profile and speed between PBC patients and healthy controls.② The level of TGF-β1 was much lower in the supernatant of BM-MS from OBC patients than health controls [(2.6±1.9)vs (8.2±6.7)ng/ml,t=-3.641,P=0.001].There were no other differences between two groups' BM-MSC.③ The super natant concentration of interlukin-10 of the third BM-MSC subculture from healthy controls was lower than that of the primary subculture [(18.5±5.0) vs (12.4±3.1) pg/ml,t=2.368,P=0.045],and that of hepatic growth factor from the second subcuhure was higher than the primary subculture [(0.21±0.07) vs (0.35±0.08) ng/ml,t=-2.874,P=0.021].There were seldom discrepancies in other cytokines between different generations of BM-MSC.Conclusion BM-MSC from PBC patients may have almost the similar characters in growth pattern and cytokines secretion as,except the TGF-β1,which was much lower than those from healthy controls.The second subculture of BM-MSC might be more suitable for the treatment to patients with PBC.

Objective To analyze the difference between primary biliary cirrhosis (PBC) and primary Sj(o)gren's syndrome (pSS) and primary biliary cirrhosis complicating with Sj(o)gren's syndrome (SS) in clinical features,anti-60 000 SSA multiple antigenic peptides (MAPs) detection rate,and explore the potential mechanisms of PBC with SS.Methods MAPs were artificially synthesized.Enzyme-linked immunosorbent assay (ELISA) was done to detect anti-MAPs antibodies in the sera of the three groups of patients.The detection rates of anti-MAPs antibodies were compared among groups and the relations of anti-MAPs antibodies with clinical features were analyzed.Chi-square test,Fisher's exact test,t test or Wilcoxon signed rank test were conducted in this study.Results There was no significant difference in clinical features,liver function tests and antibody profiles between PBC and PBC with SS.Significant difference of anti-MAP20 antibody detection rote was detected between pSS and PBC with SS groups [25％(7/28) vs 0(0/25),x2=7.201 1,P=0.007 3],and anti-MAP3,7 and 17 antibody detection rates in PBC with SS patients [4％(1/25),4％(1/25),8％(2/25)] were similar to pSS [4％(1/28),7％(2/28),7％(2/28)].The anti-MAP7 antibody and anti-MAP12 antibody detection rates were significantly higher in patients with splenomegaly than patients without splenomegaly [38％(3/8) vs 2％(2/84),38％(3/8) vs 4％(3/84); P=0.039 4,P=0.039 4],while the antiMAP17 antibody detection rate was significantly lower in patients with salivary gland injury than patients without salivary gland injury [5％(3/64) vs 39％(5/13); x2=4.431 8,P=0.035 3].Conclusion There is significant difference in the anti-MAPs detection rates among the three patients groups,and the detection rate may be higher in patients with certain clinical manifestations.

Objective To describe the immunological characteristics of refractory primary biliary cirrhosis compared with the typical patients for more than 1 year's administration of UDCA.Methods Sixty patients treated with UDCA for more than 1 year in our clinic were enrolled into this study.According to the response to UDCA by Paris criteria,patients were divided into refractory group (23 patients) and typical groups (37 patients).The recent peripheral lymphocyte subsets and cytokines of the two groups were tested and analyzed.One-way ANOVA and t test were used for statistical analysis.Results ① One-year treatment after diagnosis,there were no differences between the two groups in the distribution of peripheral lymphocytic subsets,meanwhile,the two groups had higher percentage of B cells,CD4+T cells,CD4+CD28+T cells and CD8+ CD28-T cells than healthy controls respectively.② The serum levels of IL-6 [(0.8±0.9) pg/ml vs (0.3±0.4) pg/ml] and HGF were higher in the refractory group than other groups.Conclusion During the plateau phase,refractory PBC patients have higher serum levels of IL-6 and HGF,which probably suggest that the refractory PBC patients may have severe immunologic disturbance in vivo.