The extra small virus (XSV) is a satellite virus associated with Macrobrachium rosenbergii nodavirus (MrNV) and its genome consists of two overlapping ORFs, CP17 and CP16. Here we demonstrate that CP16 is expressed from the second AUG of the CP17 gene and is not a proteinase cleavage result of CP17. We further expressed CP17 and several truncated CP17s (in which the N- or C-terminus or both was deleted), respectively, in Escherichia coli. Except for the recombinant plasmid CP17ΔC10, all recombinant plasmids expressed soluble protein which assembled into virus-like particles (VLPs), suggesting that the C-terminus is important for VLP formation.

Objective To explore the regulatory effect of astragalus polysaccharide ( APS) on immune function in immunosuppressed mice, and to establish a set of feasible indexes of immunological regulation and evaluation system of pol-ysaccharides.Methods Both Cytoxan ( CTX)-induced immunosuppressed mice and normal mice were treated with astrag-alus polysaccharide ( APS) .Spleen index, thymus index, phagocytosis rate and phagocytic index of peritoneal macrophages were assessed, and the spleen and thymus tissues were examined by histology.The percentage of CD3 +, CD4 +, and CD8 +cells in peripheral blood were determined by flow cytometry.Results APS significantly increased the thymus and spleen indexes in both immunosuppressed mice and normal mice, promoted the histological development of spleen and thy-mus in normal mice, enhanced the recovery of histological structures in the spleen and thymus in the CTX-induced immuno-suppressed mice.APS also enhanced the phagocytosis rate and phagocytic index of peritoneal macrophages in both immuno-suppressed mice and normal mice.In addition, the percentage of CD3 +and CD4 +cells,the ratio of CD4 +/CD8 +in pe-ripheral blood of both CTX-induced immunosuppressed mice and normal mice were increased while CD8 +cells were de-creased.Conclusions The results of this study suggest that astragalus polysaccharide can improve the non-specific immu-nity and cellular immunity in mice, and indicate that we established comprehensive evaluation indexes of immunoregulatory effects of polysaccharides.

The nucleocapsid protein VP15 of white spot syndrome virus (WSSV) is a basic DNA-binding protein. Three canonical bipartite nuclear localization signals (NLSs), called NLS1 (aa 11-27), NLS2 (aa 33-49) and NLS3 (44-60), have been detected in this protein, using the ScanProsite computer program. To determine the nuclear localization sequence of VP15, the full-length open reading frame, or the sequence of one of the three NLSs, was fused to the green fluorescent protein (GFP) gene, and transiently expressed in insect Sf9 cells. Transfection with full-length VP15 resulted in GFP fluorescence being distributed exclusively in the nucleus. NLS 1 alone could also direct GFP to the nucleus, but less efficiently. Neither of the other two NLSs (NLS2 and 3) was functional when expressed alone, but exhibited similar activity to NLS1 when they were expressed as a fusion peptide. Furthermore, a mutated VP15, in which the two basic amino acids (11RR12) of NLSI were changed to two alanines (11AA12), caused GFP to be localized only in the cytoplasm of Sf9 cells. These results demonstrated that VP15, as a nuclear localization protein, needs cooperation between its three NLSs, and that the two residues (11RR12) of NLS1 play a key role in transporting the protein to the nucleus.

White spot syndrome virus(WSSV), Taura syndrome virus(TSV)and Infectious hypodermal and haematopoietic necrosis virus(IHHNV)are three shrimp viruses responsible for major pandemics affecting the shrimp farming industry. Shrimps samples were collected from 12 farms in Zhejiang province, China, in 2008 and analyzed by PCR to determine the prevalence of these viruses. From the 12 sampling locations, 8 farms were positive for WSSV, 8 for IHHNV and 6 for both WSSV and IHHNV. An average percentage of 57.4% of shrimp individuals were infected with WSSV, while 49.2% were infected with IHHNV. A high prevalence of co-infection with WSSV and IHHNV among samples was detected from the following samples: Bingjiang(93.3%), liuao(66.7%), Jianshan(46.7%)and Xianxiang(46.7%). No samples exhibited evidence of infection with TSV in collected samples. This study provides comprehensive information of the prevalence of three shrimp viruses in Zhejiang and may be helpful for disease prevention control in this region.

In view of the deficiency of traditional teaching methods and teaching resources in clinical practice, our department has designed cases according to diagnosis, diagnosis basis, laboratory examination and treatment, and deploys the case based learning with PPT files according to such teaching process as case introduction, guiding discussion, summary and teaching, introspection and expansion as well as homework. Meanwhile contrast experiment was carried out. The results have showed the case based learning probably has the positive impact on improving teaching quality of clinical practice , mainly reflected in: ①contributing to the completion of the teaching outline;②contributing to the transform of theoretical knowledge from systemic structure to applied structure;③contributing to establishing the teaching model and assessment system which are more complete and standard than the traditional teaching methods; ④contributing to improving students' interest and mo-tivation;⑤contributing to improving the teaching efficiency, which makes students have more time to participate in the application and training of clinical skills.

Background:In the treatment of many gastric diseases,eradication of Helicobacter pylori( Hp)is important. With the increasing in antibiotic resistance of Hp,the eradication efficacy of conventional triple and quadruple therapy for Hp is decreasing in recent years. Aims:To assess the efficacy and safety of quadruple therapy combined with astragalus membranaceus for Hp eradication. Methods:A total of 395 Hp-infected patients with chronic gastritis/dyspepsia and peptic ulcer diagnosed by gastroscopy were enrolled,of them 185 patients were of chronic gastritis/dyspepsia group and 210 patients were of peptic ulcer group. Patients in the two groups were subdivided into trial group and control group. Patients in trial group were given quadruple therapy combined with astragalus membranaceus for 14 days,and patients in control group were given quadruple therapy only for 14 days. Hp eradication was assessed by 13 C-urea breath test 4 weeks after treatment. Results:A total of 358 patients(90. 6%)completed the study. In chronic gastritis/dyspepsia group,the eradication rate was significantly higher in trial group than in control group by PP analysis(P<0. 05),but no significant difference was found by ITT analysis(P>0. 05). In peptic ulcer group,the eradication rates by PP and ITT analysis showed no significant differences between trial group and control group(P>0. 05). In control group,the eradication rates by PP and ITT analysis were significantly higher in peptic ulcer group than in chronic gastritis/dyspepsia group( P <0. 01). In trial group,the eradication rates by PP and ITT analysis were not significantly different between chronic gastritis/dyspepsia group and peptic ulcer group( P >0. 05 ). The incidence of adverse effects was not significantly different between chronic gastritis/dyspepsia group and peptic ulcer group as well as trial group and control group( P>0. 05). Conclusions:Quadruple therapy combined with astragalus membranaceus could increase the eradication rate of Hp in chronic gastritis/dyspepsia group safely.

The nucleocapsid protein (N) is a major structural protein of coronaviruses. The N protein of bat SARS-like coronavirus (SL-CoV) has a high similarity with that of SARS-CoV. In this study, the SL-CoV N protein was expressed in Escherichia coli, purified and used as antigen. An Indirect Enzyme-Linked Immunosorbent Assay (indirect ELISA) was developed for detection of SARS- or SL-CoV infections in bat populations. The detection of 573 bat sera with this indirect ELISA demonstrated that SL-CoVs consistently circulate in Rhinilophus species, further supporting the proposal that bats are natural reservoirs of SL-CoVs. This method uses 1-2 μl of serum sample and can be used for preliminary screening of infections by SARS- or SL-CoV with a small amount of serum sample.

Objective To find the approximate error factors influence of dosing, fundamentally reduce the administra-tion of approximate error. Methods According to the three level of hospital administration approximation error of 181 cases were retrospectively analyzed. Results The following factors were caused drug similar mistakes.Administration of the approximation error incidence decreased year by year during the investigation. Medication errors and errors in the approximation, error dose administered in the proportion accounted for more than 60%. More than 90% administration of the approximation error occurs during the day. The title, length of service and equipment have important effects on reducing drug approximation error, do not comply with the work flow, violating the operating rules, because of inter-ference interrupts the continuity, the lack of communication, carelessness, memory errors, drug similarity, difference signal information system. Conclusion Specification for dug management system, the administration management as a whole,the full implementation of the"Five Rights"management.

Objective To observe and analyze the role of different subfields of medial prefrontal cortex (mPFC) in the expression course of startle reflex.Methods 24 healthy male British kind of albino guinea pigs were randomly divided into 4 groups:anterior cingulated cortex lesion ( n =6) and sham-lesion ( n =6) ( Experiment 1 ) ; prelimbic cortex lesion/joint lesion of prelimbic cortex and anterior cingulated cortex(n=6) and sham-lesion ( n =6) ( Experiment 2 ).The animals were injected lidocaine ( lesion ) or physiological saline ( sham-lesion ).Each group received paired training of conditioned stimulus( CS,a tone) and unconditioned stimulus (US,a air puff),to observe the acoustic startle reflex(ASR) change of these groups.Results As the results of experiment 1 suggested,SR rate did not change significantly after anterior cingulated cortex lesion ( time effect:F =15.421,P =0.098 ; group effect:F =14.753,P =0.084).As the results of experiment 2 suggested,SR rate did not change significantly after prelimbic cortex lesion ( time effect:F =14.975,P =0.178 ; group effect:F =18.643,P =0.089).When prelimbic cortex and anterior cingulated cortex were lesioned at the same time,SR rate declined significantly and didn ' t recover with the following training ( group effect:F =67.743,P =0.009 ).ConclusionLesions of the prelimbic cortex and anterior cingulated cortex in mPFC cause the significantly decline of acoustic startle reflex( ASR),which don' t recover with the following training.This study indicates that mPFC involves in the regulation of ASR,but the regulation mechanism needs to be discussed.

Objective To investigate the expression of AIF, CYT C, PAF-1, caspase-3, and XIAP in Sprague-Dawley rats with spontaneous mammary neoplasms.Methods One-hundred and thirty 3-4-week old SPF Spargue-Dawley rats (♀∶♂=1∶1) were fed in a specific pathogen free (SPF) breeding barrier for 60 weeks.The occurrence of spontane-ous breast tumors was recorded and histopathology was performed to identify the types of tumors.The rats were divided into 3 groups:rats with normal breast tissue ( group I) , with benign tumors ( group II) and with malignant tumors ( group III) . The expression of AIF, CYT C, APAF-1, caspase-3 and XIAP proteins and mRNAs were detected by immunhistochemistry ( IHC) and RT-PCR assay.Results Among these 130 SD rats, 14 rats were observed having spontaneous mammary neo-plasms with the incidence rate of 10.77%(14/130).In these neoplasm cases, 7 cases were mammary fibroadenomas, 7 cases of breast carcinoma, both with an incidence rate of 5.38%.Immunohistochemistry showed that, compared with the group I, the positive expressions of AIF, APAF-1, caspase-3 were decreased significantly (P<0.01), and the CYT C and XIAP expressions were significantly increased in the group II.The positive expression of all genes except XIAP was de-creased in the group III(P<0.01).Compared with the group II, APAF-1 and XIAP were significantly higher in the group III (P<0.01), and the positive expression of AIF, Cyt C, and caspases-3 were significantly decreased (P<0.01).In the results of RT-PCR assay, except APAF-1 which showed significant correlation with the results of immunohistochemistry ( P<0.05 ) , all the others showed an extremely significant correlation with immunohistochemical results ( P <0.01 ) . Conclusions Mammary tumors are most common spontaneous neoplasms in SD rats.Abnormal expression of mitochondrial apoptotic pathway-related factors AIF, CytC, APAF-1, caspase-3, and XIAP are correlated with the carcinogenesis and de-velopment of breast tumors.