A novel method for DNA detection was developed based on the excellent fluorescence properties of cationic conjugated polymer ( CCP)and the target DNA enrichment,separation function of nanoparticles. First,the quencher-labeled DNA capture probes were modified on the surface of Au nanoparticles,and complementary DNA strands were captured. Second,S1 nuclease was added,and the capture probes that had not captured the complementary DNA were removed from the nanoparticles.Finally,the complementary double-stranded DNA was cut by Dnase I,the quenchers were dissociated from nanoparticle and the fluorescence of CCP was quenched by means of combination of quenchers and CCP.The results showed that this method is specific.In the range of 5. 0 -40 nmol/L,the concentration of target DNA was proportional to the fluorescence quenching and the detection limit was 3.7 nmol/L(S/N = 3).

Purpose To investigate the expression of tumor buds and its relationship with clinicopathological characteristics in endome-trial carcinoma. Methods 47 cases of endometrial carcinoma were observed and analyzed by means of clinicopathologic data and im-munohistochemical staining. The connections between tumor buds and clinicopathologic parameters were studied by statistics. Results Of the 47 endometrial carcinoma cases, tumor buds were seen in 18 cases. Tumor buds were correlated with histological grade, lymph node metastases, vascular invasion, Ki-67 index and survival. There were no associations between tumor buds with age, tumor size, pTNM stage or myometrial invasion depth. Immunohistochemistry demonstrated that the level of N-cadherin, vimentin in bud cells was higher than normal carcinoma cells while the level of CK(AE1/AE3),β-catenin and E-cadherin got the opposite results. Conclu-sion Tumor buds may play an important role in the progression of endometrial carcinoma. The immunohistochemical features of bud cells indicated that tumor buds may be a key step in epithelial-mesenchymal transition.

Objective To develop an antigen retrieval method for detection of human mammaglobin ( hMAM) immuno-histochemcal staining in old paraffin-embedded specimens .Methods The tissue sections in test group were put into dis-tilled water after deparaffinization and then moved into citric acid buffer ( pH 3.5) for 10-15 min.The other two meth-ods,microwave method and high pressure cooker method ,were compared as control groups at the same time .Finally, immu-nohistochemistry SP method was used to check the antibody in the sections .Results The color appearance in the test group (pH 3.5 citric solution) was better than that of microwave oven and high pressure cooker groups .In the test group, tissue sections were not easily cast off from the slices .Conclusion In this study,we have established a new and simple antigen retrieval method which will contribute to immunohistochemistry technology .

Objective To construct prokaryotic expression vectors for glutamate dehydrogenase(GDH)of Clostridium difficile(CD),and express recombinant GDH in Escherichia coli,and identify its antigenicityed.Methods The entire gene of GDH was cloned from ATCC43255 genome DNA.The recombinant antigens were expressed in E.coli with IPTG induction and purified by Ni-NATBeads.The antigenicity was detected using CD Qick Chek Complete dual-antigen EIA. Results Prokaryotic expression vectors of CD GDH were constructed successfully.The antigen could be identified by specific anti-GDH antibodies.Conclusion The GDH antigen can be used to prepare corresponding antibodies,which facilitate the development of immunoassay for CD GDH.

Objective To analysis the positive rates of glutamic acid decarboxylase autoantibody (GADA)and zinc transporter 8 autoantibody (ZnT8A)in newly diagnosed type 2 diabetes patients.Methods GADA and ZnT8A were detected in 101 ca-ses of newly diagnosed type 2 diabetes mellitus patients using ELISA.Results The positive rate of GADA was 21.78%,the positive rate of ZnT8A was 17.82%,and the common positive rate of GADA and ZnT8A was 8.91%.There were no corre-lations between GADA or ZnT8A autoantibodies and the patient’s sex (t=-0.724,-0.550;0.903,1.359,P >0.05),age (t=-0.724,-0.550;0.903,1.359,P >0.05),blood glucose (r=0.290,0.110;-0.264,-0.047,P >0.05),cholesterol (r=-0.047,0.004;0.154,-0.138,P >0.05),triglyceride (r=-0.092,-0.054;-0.217,-0.023,P >0.05),and low density lipoprotein (r= - 0.045,- 0.027;0.202,- 0.025,P > 0.05).Conclusion It should be screened autoantibodies timely for newly diagnosed type 2 diabetic patients in order to diagnosis the Latent autoimmune diabetes in adults early.

Objective:To investigate the effects of abdominal Tuina(Chinese therapeutic massage)on behavioral function,5-hydroxytryptamine 1A receptor(5-HT1AR),and synapsin-1(Syn1)in neonatal rats with hypoxic-ischemic brain injuries(HIBI). Methods:Forty healthy neonatal rats,born of 5 specific pathogen-free healthy pregnant rats,were randomly divided into a group for modeling(n=28)and a sham operation group(n=12)on the 7th day of birth.In the group for modeling,24 neonatal rats with HIBI successfully established by the Rice method were randomly divided into a model group(n=12)and an abdominal Tuina group(n=12).The abdominal Tuina group was given abdominal Tuina for 28 d from 24 h after modeling,and the other groups were put under the same conditions but without any treatments.Rats in each group were subjected to suspension tests on the 7th,14th,21st,and 28th days of intervention.After the intervention,the rat hippocampal tissue was collected and stained with hematoxylin-eosin to observe the pathological changes in the rat hippocampal CA1 region.The 5-HT1AR expression in rat hippocampal CA1 region was detected by immune-histochemistry.The Syn1 expression in rat hippocampus was measured by Western blotting method. Results:The cells were disordered,and edema and necrosis appeared in the hippocampal CA1 region of the model group.Cell arrangement was clear,and edema was improved obviously in the hippocampal CA1 region of the abdominal Tuina group.Compared with the sham operation group,the suspension test scores,the number of 5-HT1AR positive cells,and Syn1 protein expression in the hippocampus decreased significantly in the model group after 21 d and 28 d of interventions(P<0.05).Compared with the model group,the suspension test scores,the number of 5-HT1AR positive cells,and Syn1 protein expression increased significantly in the abdominal Tuina group after 21 d and 28 d of interventions(P<0.05). Conclusion:Abdominal Tuina improves the behavioral function of upper limbs and up-regulates the expression levels of 5-HT1AR and Syn1 in the hippocampus of neonatal HIBI rats.

Objective To develop the technique to detect total core antigen of HCV(Total HCV-cAg) by Enzyme-Linked Immu-nosorbent Assay (ELISA) and apply it for clinical diagnosis. Methods 201 serum samples with anti-HCV antibody were detected total HCV-cAg after pre - treating the samples, then the sensitivity of results were compared with HCV RNA tests. Among them, 176 cases was determined by FQ-PCR, and 25 cases by RT-PCR for HCV-RNA. Results HCV RNA was found in sera from 88 of 201 samples (43.8%). Total HCV-cAg was positive in 71 (35.3%) of 201 samples . There was no significant difference between the detection rate of HCV RNA by PCR and total HCV-cAg by ELISA. Conclusion Detection of total core antigen of HCV is suitable to be used as to diagnose HCV in clinic.

Objective To obtain different fragments of human carboxypeptidase H,and evaluate the diagnostic application of the recombination carboxypeptidase H in detecting autoantibody.Methods The coding gene of carboxypeptidase H was ob-tained by RT-PCR.The corresponding prokaryotic expression vectors were constructed and transformed into E.coli to in-duce the expression of the recombination different fragments of carboxypeptidase H.Using these antigen fragments as the coating antigens,the enzyme-linked immunosorbent assay (ELISA)was established for the detection of carboxypeptidase H autoantibody in 95 newly diagnosed type 2 diabetes patients.Results Three fragments of human carboxypeptidase H were obtained,in which the 42~476aa fragment antigen was ideal one.Using the full-length carboxypeptidase H as coating anti-gen,the positive rate of carboxypeptidase H autoantibody was 8.42%.Conclusion Because of the favorable antigenicity,the 42~476aa fragment antigen of carboxypeptidase H could be the candidate antigen for discrimination and diagnosis of latent autoimmune diabetes in adults.

Objective To explore the effect of high-frequency repetitive transcranial magnetic stimulation (rTMS) on activity in the intact motor cortex controlling the suprahyoid muscles and thus on dysphagia after an unilateral stroke.Methods Forty patients suffering dysphagia more than two weeks after a unilateral cerebral stroke were randomly divided into an experimental group and a control group,each of 20.Both groups were given traditional swallowing rehabilitation training,while the experimental group was additionally provided with 5 Hz rTMS for two weeks.Before and after the treatment,all of the patients were characterized using X-ray fluoroscopy,video fluoroscopic swallowing study (VFSS) and surface electromyography,and their swallowing was evaluated using a standardized swallowing assessment (SSA) and a penetration-aspiration scale (PAS).Results After the treatment,significant improvement was observed in both groups in the average swallowing time and in the maximum amplitude of sEMG,as well as the average SSA,PAS and VFSS scores (P＜0.05).The average values in the experimental group were in all cases significantly better than the control group's averages (P＜0.05).Conclusion Applying rTMS at 5 Hz to the motor cortex of the contralateral hemisphere controlling the suprahyoid muscles can effectively improve unilateral-hemisphere dysphagia after stroke.