The antigen of hemorrhagic fever with renal syndrome (HFRS) virus in the infected suckling mouse brains was purified by a combined method—Protamin sulfate sedimentation and sucrose cushion uhracentrifugation. The Purified antigen was labeled with horseradish peroxidase (HRP). A kind of IgM antibody capture ELISA that used HRP—labeled antigen of HFRS Virus, i.e, direct ELISA, was successfully established for the detection of specific IgM antibody in sera from patients with HFRS. Compared with IgM antibody capture ELISA that used HRP—labeled IgG antibody to HFRS virus ,direct ELISA was similar in sensitivity to it,but direct ELISA could completely avoid the interference caused by rheumatoid factor (RF) as well as could reduce one step immune reaction. 87 serum samples from patients with HFRS(diagnosed clinically) in various stages, including 37 from Patients with HFRS in early stage(from 1 to 5 days after the onset of HFRS)were detected by direct ELISA . The positive rates of specific IgM antibody were 96.5% and 91.8%, respectively. We think that direct ELISA that uses HRP—labeled antigen provides a more specific,simpler and faster method for early di—agnosis of HFRS.

Objective To study the histological structures of retinotectal system in zebrafish. Methods The structures of tectum and retina were examined histologically and the total number and diameter of the optic nerve fibers were studied quantitatively. Results The retina includes 10 layers, and the tectum is differentiated into 6 layers. The mean thickness of the tectum is about 219\^7?8\^3?m. The optic nerve of zebrafish has approximately 78?960 myelinated fibers. The mean of the outer diameter (D) of the myelinated fibers was 0\^50?0\^18?m, while the mean of the inner diameter (d) is 0\^35?0\^14?m. Virtually all axons within optic nerves are myelinated.Conclusion\ The results not only accord with the typical pattern of retina and tectum in vertebrates, but also reflect the characteristic of diurnal activity and good vision in zebrafish.\;[

Objective To explore the mechanism of azithromycin (AZM) for inhibiting the proliferation of rat airway smooth muscle cells (ASMCs).Methods Thirty Sprague-Dawley (SD) rats were divided into the control group,asthma model group and AZM group.The rat model of asthma was established by ovalbumin (OVA) sensitization and stimulation in vitro.The airway related parameters of rat lung tissue were determined by using the medical image analysis system.Primary passage ASMCs were isolated and cultured using the tissue-sticking method,and the vascular endothelial growth factor (VEGF) overexpression vector or tumor necrosis factor receptor-associated factor 6 (TRAF6) overexpression vector was transfected into ASMCs in the AZM group.The protein levels of VEGF,NF-κB p65 and TRAF6 were detected by Western blotting,and the proliferation of ASMCs was evaluated by CCK-8 kit.Results AZM significantly inhibited the increase of thickness of total airway wall,thickness of inner airway wall and thickness of airway smooth muscle layer in asthma rats (P<0.05),also significantly inhibited the proliferation of ASMCs in the asthma model group (P<0.05).AZM significantly inhibited the protein expression of VEGF and NF-κB p65 induced by asthma (P<0.05),and the overexpression of VEGF significantly reduced the inhibiting effects of AZM on proliferation of ASMCs (P<0.05).AZM significantly inhibited the high expression of TRAF6 induced by asthma (P<0.05),and the overexpression of TRAF6 significantly reduced the inhibiting effects of AZM on expression of VEGF and NF-κB p65 as well as proliferation of ASMCs (P<0.05).Conclusion AZM can suppress the proliferation of ASMCs,its partial mechanism may be realized through inhibiting TRAF6/NF-κB/VEGF signaling pathway.

Objective To describe the characteristic MR appearances in hepatocellular carcinoma after radiofrequency ablation (RFA) during follow-up studies. Methods MR images of 110 patients with hepatocellular carcinoma after RFA ,which were categorized into 3 MR examination intervals ( ≤48 h, 1-6 m, ＞6 m), were analyzed retrospectively. The sequential changes of MR images were assessed and compared using Chi-square test. Results All RFA areas of 110 patients typically exhibited hyperintensity on GRE-T1 WI and hypointensity on TSE-T2 WI with fat suppression within 48 h after ablation. Subsequently,the signal intensities of the RFA areas decreased on GRE-T1 WI which showed significant differences after 6 months( P ＜0. 015 ). On GRE-T1 WI , RFA lesions were hyperintense in 72, iso-or hypo-intense in 4 at 1-6 months and hyperintense in 60, iso-or hypo-intense in 17 after 6 months. On TSE-T2 WI, 65 RFA lesions were hypointense, 11 isointense at 1-6 months and 47 were hypointense, 30 isointense after 6 months. The increased intensity on TSE-T2 WI showed significant differences after 6 months( P ＜ 0. 015 ).On contrast-enhanced MR images, RFA lesions showed peripheral rim of enhancement within 48 h and exhibited a tendency to show no enhancement ( n = 37 at 16 months and n = 63 after 6 months ) with significant differences between MR examination intervals( P ＜ 0. 015 ). In 6 patients with residual tumor or local tumor progression, nodular lesions were found at the periphery of the RFA. These nodular lesions showed moderate to marked enhancement as well as hypointense signal on GRE-T1WI, and moderate hyperintense signal on TSE-T2WI. Conclusion MR imaging could depict the characteristics of coagulative necrosis of tumor, residual tumor or local tumor progression in patients with hepatocellular carcinoma after RFA therapy.

Objective To discuss the diagnostic value of CT and MRI for primary anorectal malignant melanoma(AMM).Methods 5 patients(2 males and 3 females)with AMM confirmed pathologically were analyzed retrospectively,which were examined by CT(n=4)and 1.5T MRI(n=1).Results AMM presented as bulky intraluminal fungating masses without intestinal obstruction(n=4)and irregular rectal wall thickening(n=1).2 cases had perirectal fat infiltration and extended to the presacral space in one case,4 cases had perirectal enlarged lymph nodes and larger than 3 cm in diameter in one case.Nonenhanced CT showed a hypodense lesion and contrast-enhanced CT showed slight enhancement(n=1)and mild enhancement(n=3).The tumor scanned by MRI depicted high intensity on T1WI and low intensity on T2WI,so did the enlarged lymph nodes and bone metastasis.Fat-saturation T1WI significantly improved the extention of tumor and detection of metastasis.Conclusion Bulky intraluminal fungating masses without intestinal obstruction,perirectal fat infiltration and enlarged lymph nodes may be helpful to diagnose AMM.MRI is especially useful in differentiating melanotic AMM from other rectal tumors.

Objective To investigate the relationship between inflammatory bowel disease (IBD) activity and choroidal thickness,and evaluate the utility of a choroidal thickness measurement in assessing IBD activity.Methods A total of 100 eyes of 50 patients of IBD with different disease activity,including 23 patients of ulcerative colitis,27 patients of Crohn's disease (CD).Ninety-six eyes of 48 healthy volunteers were recruited as control group.Choroidal thickness was measured using enhanced depth imaging(EDI)optical coherence tomography.Results Compared with the subfoveal choroidal thickness (294.37 ± 35.04) μm in healthy volunteers,the subfoveal choroidal thickness (349.28 ± 76.57) μm in UC patients with severe disease activity,the subfoveal choroidal thickness (326.71 ± 59.71) μm and (354.24 ± 66.34) μm,respectively,in CD patients with moderate and severe disease activity were found to be increased significantly (all P ＜ 0.05).Conclusion Choroidal thickness should be considered as a potential marker to assess the disease activity in patients with IBD,especially in patients with CD.

Objective To investigate the FECH gene mutation in a Chinese family with erythropoietic protoporphyria, to explore the relationship between gene mutation and clinical manifestations so as to estab-lish a basis for the genetic diagnosis and treatment of erythropoietic protoporphyria. Methods Clinical data on a Chinese family with typical EPP was collected. Peripheral blood was obtained from patients, unaffected individuals in the family and 50 unrelated human controls. Genomic DNA was extracted and PCR was per-formed to amplify the whole coding regions (exons 1 to 11) of FECH gene and their flanking intron sequences followed by direct sequencing to detect possible mutations. Results Based on clinical symptom and por-phyrin levels, a diagnosis of erythropoietic protoporphyria was made in 3 family members. DNA fragments of expected size were amplified by PCR. Gene sequencing revealed a heterozygons mutation (IVS1 + 1G >C) in intron 1 of FECH gene in the proband, his sister and father, but not in unaffected family members or unrelated human controls. Also, an IVS1-23C/T polymorphism associated with low expression alleles was observed in intron 1 of FECH gene of the proband, his sister and mother. Conclusions A novel mutation in the donor splice site of intron 1 of FECH gene is first reported in a Chinese family with EPP; this muta-tion may lead to a deficiency of FECH gene and serve as a molecular basis of development of erythropoietic protoporphyria.

Objective To detect anti-HCV in serum of hepatic disease patients by performing the confirmatory test, and further to confirm HCV infection. Methods Two recombinant immunoblot assays (CWT and CHIRON RIBA HCV 3.0 Strip Immunoblot Assay) were used respectively to detect anti-HCV in 477 human serum samples, which comprised 350 HCV-infected patients' specimens, 7 none-A none-E hepatitis specimens, 30 HBV-infected patients' specimens, 30 hepatitis E virus infected patients'specimens, and 60 specimens drawn from blood donors. The latter three groups served as controls. Results A total of 120 control non-HCV-infected patients' specimens were negative when tested by both assays. Among 350 HCV-infected patients, 341 were positive and 9 were indeterminated by CWT assay; 343 were positive and 7 were indeterminated by CHIRON RIBA HCV 3. 0 SIA. Seven none-A none-E hepatitis specimens tested by both assays turned out to be 2 positive, 4 negative and 1 indeterminate. The consistency rate of these two assays was 99. 16% (Kappa=0.98). Conclusion CWT assay is highly coherent with CHIRON RIBA HCV 3.0 SIA assay in the methodology of anti-HCV antibody detection, which can be applied in the determination of HCV infection among none-A none-E hepatitis patients.

Objective To summarize the clinical experience of treating long bone defect with vascularized fibular graft.Methods From January,2008 to January,2015,31 cases of long bone defect were treated with vascularized fibula composite or not composite tissue flap graft.The length of transplanted fibula was 9-20 cm,and the flap area was 5 cm×3 cm to 21 cm×14 cm.All patients were followed up regularly.Limb function was assessed 12 months after surgery.Results Thirty-one cases of vascularized fibular flap survived after surgery.Thirty patients were followed up for 1.5 to 6 years (average,2.5 years).One patient was lost to follow-up.The bone defects of patients followed up were healed.There was one case fracture occurred for trauma,was treated with plaster cast for 6 months and healed.The transplanted fibular thickened for 1.3 to 2.5 years(average,1.6 years).Conclusion Vascularized fibular graft can reconstruct long bone defect for single use and shorten the duration of treatment with a good limb function.For cases combined soft tissue defect,vascularized fibula composite tissue flap can be applied to repair at the meantime.

Objective To analyze the computed tomography(CT)and magnetic resonance imaging (MRI)appearances of primary clear cell carcinoma of the liver (PCCCL)and evaluate the value in the diagnosis of the disease.Methods CT and MR images of 38 patients of pathologically confirmed PCCCL were evaluated retrospectively.Twenty-six patients underwent CT,23 underwent MRI, and 1 1 underwent both CT and MRI.Results In pre-contrast CT scanning,24 PCCCLs appeared hypodense and 2 hyperdense.As for MRI 1 9 of the 23 PCCCLs were hypointense and 4 were iso-hypointense on T1 WI.While on T2 WI,22 cases were heterogeneously hyperintense,and 1was iso-hypointense.On the arterial phase of CT/MRI,all cases presented intense enhancement,and on the portal venous phase,35 cases(35/38,92.1%)were relatively hypodense/hypointense and 3 were slightly hyperdense/hyperintense.Among the tumors larger than 3 cm(n=22),nodular enhancement pattern was found in 14 cases(63.6%,14/22).The capsular rim en-hancement was demonstrated in 26 cases.Conclusion PCCCL showed similar dynamic enhancement pattern as common hepatocellu-lar carcinoma,but also depicted specific imaging features.