Objective To study the distribution and tumor targeting property of anti-human prostate specific membrane antigen(PSMA) monoclonal antibody Ed-5 labeled with ~ 131 I in nude mice bearing human prostate cancer. Methods The nude mice xenografted models of human prostate cancer were established. Anti-human PSMA monoclonal antibody Ed-5 was labeled with ~ 131 I. ~ 131 I-Ed-5 was injected through the tail vein of nude mice bearing human prostate cancer. r-ray scintigraphy was performed at 24,48,96 and 144 hours after injection. And then the mouse were killed, the radioactivity ratio of tumor and non-tumor (T/NT), percentage of injected dosage of every gram tissue (%ID/g) were measured, and the bio-distribution of the labeled antibody was determined. Results The tumor imaging was clearly visualized at 96 hours after ~ 131 I-Ed-5 injection, and the tumor brim was clearer along with time prolonging. Radioactivity assembled in the tumors 24 to 144 hours after ~ 131 I-Ed-5 injection. The tumor's %ID/g at 96 hours after injection was 32.30, the radioactivity ratio of tumor and various tissues reached a maximum, and that of tumor/muscle was 7.08. Conclusion Ed-5 exhibited excellent immunoreactivity and tumor targeting property, and had application potential in targeting diagnosis and therapy of prostate cancer.

Objective To understand the structures and functions of enterovirus 71 (EV71)VP1 gene and its encoded protein using bioinformatics method, so as to direct studies of its biological function. Methods VP1 gene and its encoded protein of EV71 2008-GZCH07 strain and other representative EV71 strains were analyzed by online analysis at bioinformatics websites and software packages. Multi-sequence homological alignment and phylogenetic analysis were performed.Physicochemical characteristics, secondary structure, homology modeling of tertiary structure,enzymological characteristics, antigenic epitope of VP1 gene encoded protein were predicted. Results The homology of EV71 2008-GZCH07 strain was highest (97% and 98%) with ZJ001 strain, and lowest with human coxsackievirus A16. The homology of EV71 2008-GZCH07 strain and EV71 types A,B,C was 86%-98%. Phylogenetic analysis demonstrated that 2008-GZCH07 stain was close to ZJ001 and BJ08-Z025-5 stains, which belonged to C4 subtype. In VP1 encoded proteins of EV71 types A,B,C, the relationship between 2008-GZCH07 and EV71-B, EV71-C was closer than EV71-A.The whole length of VP1 gene was 510 bp, with open reading frame (ORF) located at 116-510 bp region,and it encoded 132 amino acids with isoelectric point of 4.39.The protein was rich of a-helix and random coilon without transmembrane regions, and contained 5 high hydrophobic regions and belonged to extracellular protein. The homology modeling of tertiary structure showed that the region was on the surface of protein and formed a binding loop. There was 5 antigen epitopes. And 7 key catalytic sites were located at or close to the loop. Conclusions EV71-VP1 encoded protein contains many phosphorylation sites, with many biological function sites and antigenic epitope regions, which might be a potential target antigen for immunodiagnosis, anti-schistosome drug and vaccine development, and would be basis of further study of diagnosis, treatment and prevention of EV71 infection.

Objective In order to assess the value of Echinoccocus granulosus cyclophilin A (EgypA) in immune diagnosis,this novel gene was cloned and expressed.Methods By screening the EST library,the coding region of EgCypA was identified,and the PCR primers were designed based on this sequence.Bioinformatic tools were used to deduce the amino acid sequence of EgCypA and analyzed its biological characteristics.EgCypA was amplified from E.granulosus cDNA library by using PCR.Then,it was cloned into the prokaryotic expression vector pET28a and transformed into E.coli BL21.The recombinant protein was expressed in E.coli BL21 after IPTG induction.The immunogenicity of EgCypA was evaluated by Western blot using the Echinoccocus granulosus and other parasites infected animals' sera.Results Results of SDS-PAGE electrophoresis show that the recombinant BL21 expressed 18 400-25 000 protein,which was identical with the molecular weight calculated by bioinformatic analysis.Western blot shows that the recombinant protein only reacted with its immune serum and E.granulosus cystic fluid immune serum,and EgCypA immune serum could react with the excretion and secretion antigen from E.granulosus protoscolexes,and no cross-reaction between EgCypA and sera from other parasites infected animals.Conclusions Cloning and expression of EgCypA are successful.EgCypA has good immunologic activity and could be a candidate molecular for immune diagnosis of echinococcosis in early stage.

AIM: To observe the effect of (IFN-?1b) on morphology and the expression of connective tissue growth factor (CTGF) in hepatic stellate cells (HSC). METHODS: HSC were cultured in vitro, and were treated with (IFN-?1b) and TGF-?_1. The changes of HSC on morphology were observed, and the expression of CTGF in HSC was assessed using reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: Phenomenon of apoptosis were observed in HSC treated with IFN-?1b. The expression of CTGF in HSC was decreased after treatment with (IFN-?1b) in control and TGF-?1-triggered groups. CONCLUSION: (IFN-?1b) is able to induce the apoptosis and suppress the expression of CTGF in HSC.

The means of survival of genomic dsRNA of reoviruses from dsRNA-triggered and Dicer-initiated RNAi pathway remains to be defined.The present study aimed to investigate the effect of Grass carp reovirus (GCRV) replication on the RNAi pathway of grass carp kidney cells (CIK).The dsRNA-triggered RNAi pathway was demonstrated unimpaired in CIK cells through RNAi assay.GCRV-specific siRNA was generated in CIK cells transfected with purified GCRV genomic dsRNA in Northern blot analysis; while in GCRV-infected CIK cells,no GCRV-specific siRNA could be detected.Infection and transfection experiments further indicated that replication of GCRV correlated with the increased transcription level of the Dicer gene and functional inhibition of in vitro synthesized egfp-siRNA in silencing the EGFP reporter gene.These data demonstrated that although only the genomic dsRNA of GCRV was sensitive to the cellular RNAi pathway,unidentified RNAi suppressor protein(s) might contribute to the survival of the viral genome and efficient viral replication.

Objective To analyze the immunological characteristics of Trichinella spiralis secretory antigen P53 and to evaluate its value in diagnosis of trichinellosis. Methods An open read frame of secretory antigen P53 was cloned from Trichinella spiralis by reverse transcriptasepolymerase chain reaction (RT-PCR) and then sequenced. Bioinformatics analysis was performed to search for its homologues in other helminths and predict its potential linear B cell epitopes and T cell epitopes. The sequence coding mature peptide was inserted into prokaryotic expression vector pET28a(+) and the purified recombinant product was identified by Western blot using serum samples of patients infected with Trichinella spiralis or other helminth. Results Bioinformaties analysis results showed that there was no P53 homologue in other helminths, which indicated that there were many linear B cell epitopes and T cell epitopes in TsP53. The recombinant P53 antigen only reacted with the serum samples of patients infected with Trichinella spiralis without any cross-reaction with the serum of patients infected with other helminths. Conclusion P53 has strong immunogenicity and immunoreactivity, which may be a promising candidate for developing Trichinella spiralis specific diagnostic method.

Objective To study the expression of tumor type M2 pyruvate kinase(tumor M2-PK) in nonsmall cell lung cancer (NSCLC) and their correlation with treatment response and prognosis.Methods The concentration of tumor M2-PK in plasma was detected by ELISA in 106 healthy controls and 83 NSCLC patients.The patients were followed for 24 months.Results The patients after surgical operation showed marked reduction in plasma tumor M2-PK level(13.5 U/ml vs 25.4 U/ml,P

Objective To study the genetic diversity in Fujian Sanming Sarcandra Glabra from different geographical provenances;To construct genetic relationship diagram.Methods DNA in leaves was extracted by CTAB. Analysis and evaluation of DNA molecular level of 45 samples of different geographical provenances were conducted by ISSR method. POPgen32 software was used to calculate the genetic diversity and establish gene trees. NTSYS software was used to carry out cluster analysis.Results Six ISSR primers amplified 630 bands. Genetic diversity analysis showed that the average effective number of alleles of 45 varieties was 1.620 2;the average Nei's genetic diversity index was 0.364 5;the average Shannon information index was 0.543 2. Each point had different levels of genetic diversity. Nei's genetic diversity index of the maximum value was 0.497 2, and the minimum value was 0.107 8;Maximum Shannon information index was 0.690 3, and the minimum value was 0.219 2. Cluster analysis results showed that 45 varieties and 14 loci band data were the primitive matrix. 630 genetic similarity coefficients between two different species were obtained. The maximum similarity coefficient among different groups was 1.0, and the minimum was 0.516.Conclusion Different varieties of Fujian Sanming Sarcandra Glabra exist abundant genetic variation and has the molecular basis of abundant species. Using 0.610 as the threshold value can divide Sarcandra Glabra from 45 different geographical provenances into 6 groups. The genetic distance and geographical distance was not related.

Objective To investigate the clinical characteristics and surgical outcomes of pancreatic acinar cell carcinoma (PACC).Methods The clinical and follow up data of PACC patients treated with surgery in Zhongshan Hospital of Fudan University between 1999 and 2012 were analyzed retrospectively.Results A total of 16 PACC were identified including 13 male and 3 female patients.The age of the patients ranged from 38 to 71 with an average of 57 years old.Six patients presented as abdominal pain,while low back pain in 3 patients,abdominal distention in 2 patients,emaciation in 2 patients,jaundice in 2 patients and melena in 1 patient.Elevated CA19-9 level was observed in 8 patients and 2 patients had elevated serum CEA.The tumors were located in the uncinate process in 1 patient,head in 9,body and tail in 6.The superior mesenteric vein was invaded in 2 cases and 1 patient had hepatic artery invasion.The tumor invaded both the celiac trunk and splenic artery in 1 patient.One patient had just undergone intra-operative needle aspiration biopsy due to unresectable tumor.All the other 15 patients underwent surgical excision with R0 resection.Among the 10 patients received pancreaticoduodenectomy,2 had superior mesenteric vein resection and replacement and 1 had hepatic artery resection.Five patients underwent distal pancreatectomy without spleen preservation.The mean size of these tumors was 5.7 cm × 4.6 cm,12 cases had a surrounding envelope,while lymphatic metastasis was observed in 8 cases.The follow up data of 15 patients were collected and the median postoperative survival was 21 months,and the survival rate of 1,3,5 year was 71.4％,28.6％,7.1％.Conclusions Pre-operative diagnosis of PACC is extremely difficult due to lack of specific clinical features and lab tests.Surgery is the first line treatment for PACC,and the prognosis of PACC is better than that of pancreatic ductal adenocarcinoma.

Objective: To investigate the automatic implicit attitude and explicit stigma of mental illness, and the intervention effectiveness on the stigma of mental illness in college students. Methods: Single category implicit association test(SC-IAT) and questionnaire survey were used to study the degree of implicit and explicit mental illness stigma among 30 college students of Peking University, and investigated the effect of two strategies, education and contact.Data were analyzed by SPSS 22.0 with t-test and variance analysis. Results: The D values of SC-IAT in the three dimensions of cognitive evaluation, emotional response and behavioral tendency of college students were 0.29±0.37, 0.24±0.43, and 0.10±0.43.There was a statistically significant difference between the cognitive evaluation dimension, the emotional response dimension and 0(t=4.069, P<0.01; t=2.925, P<0.01). There was no statistically significant difference between the implicit stigma and the 0 in the behavioral tendency dimension(t=1.423, P=0.167). In the explicit stigma, the mercy dimension score(1.94±0.61) was lower than the median, and the difference was statistically significant(t=-12.286, P<0.01), while the scores of anger dimension(4.62±0.44) and fear dimension (4.55±0.31) were higher than the median(t=13.243, 3.200, both P<0.01). There were no significant differences in the values of implicit and external stigma among pre-intervention, education intervention and after contact(P>0.05). The correct rates of questions "Which of the following situations do you think are considered mentally unhealthy? " and "Which of the following disorders are mental illnesses" (42±13)%, (54±13)%) increased after receiving educational intervention ((49±12)%, (64±13)%), the difference was statistically significant (F=3.682, P<0.05; F=3.576, P<0.05). Conclusion College students have implicit mental illness stigma, educational and contact intervention strategies can help improve college students' understanding of relevant knowledge, and cannot improve the level of implicit stigma of college students on mental illness.