A label-free activatable biosensor for detection of exosomal microRNAs based on DNA-AgNCs and hairpin type nucleic acid probes.

Exosomal microRNA (miRNA) is a potential biomarker for cancer diagnosis, metastasis, and treatment. detection of exosomal miRNA is an attractive option due to its simplicity and high accuracy. However, exosomal miRNA detection has encountered challenges because of the low target abundance of targets and limited probe permeability.

Investigation on the co-infections of with PRRSV, CSFV or PCV-2 in swine in part of China.

The objective of the present investigation was to estimate the prevalence of infection and co-infection with porcine reproductive and respiratory syndrome virus (PRRSV), classical swine fever virus (CSFV) and porcine circovirus type 2 (PCV-2) in pigs in China. A total of 372 tissues or serum samples collected from pigs distributed in 9 provinces/municipalities of China during the period from February 2011 to November 2012 were assayed for antigens and antibodies using enzyme linked immunosorbent assay (ELISA) technique, while the PCR was designed for the detection of the PRRSV, CSFV and PCV-2, respectively. The total positive rate of PRSSV, CSFV and PCV-2 was 9.

High prevalence of torque teno sus virus in China and genetic diversity of the 5' non-coding region.

Members of the family Anelloviridae are emerging circular DNA viruses infecting many species of vertebrates including pigs. To date, members of two distinct genera, Iotatorquevirus, including torque teno sus virus 1a and torque teno sus virus 1b (TTSuV1a and TTSuV1b), and Kappatorquevirus, including torque teno sus virus k2a and torque teno sus virus k2b (TTSuVk2a and TTSuVk2b), have been identified in domestic pigs and wild boars. The goal of this study was to evaluate the prevalence and genetic diversity of these viruses based on 5' non-coding genes in Chinese swine herds experiencing clinical symptoms.

High prevalence of bovine viral diarrhea virus 1 in Chinese swine herds.

Nested RT-PCR was used to investigate bovine viral diarrhea virus in 511 specimens collected from Chinese pigs exhibiting clinical symptoms between 2007 and 2010. Of these, 137 samples were BVDV-positive and the BVDV prevalence rate was 23.1% (9/39) in 2007, 27.

[Study on liver injury of Oncomelania hupensis caused by Eomecon chinanthe sanguinarine].

Objective: To analyze the effects of Eomecon chinanthe sanguinarine (SAN) on glucogen, enzyme activity and lipid peroxidation of Oncomelania hupensis liver so as to explore the mechanism of SAN against Oncomelania hupensis.

Methods: SAN was extracted and purified from the dry powder of Eomecon chionantha. Oncomelania hupensis were immersed in 5 mg/L sanguinarine (50 Oncomelania hupensis per 500 ml solution) or clean water at 25 degrees C for 36 h, the livers were isolated from live snails.

Co-existence of multiple strains of porcine circovirus type 2 in the same pig from China.

Pigs are often co-infected by different viral strains from the same virus. Up to now, there are few reports about co-existence of different porcine circovirus type 2 (PCV2) strains in China. The aim of this study was to evaluate it in Chinese swine herds.

The RNA profile of porcine parvovirus 4, a boca-like virus, is unique among the parvoviruses.

PPV4 transcribes its genome from a single promoter, and the RNAs are generated via alternate splicing coupled with alternate polyadenylation, a strategy similar to that of the bocaviruses; however, several differences were detected. The PPV4 ORF1 codes for four NS proteins, while the bocavirus ORF1 codes for 1-3 NS proteins. Whereas the VP1/VP2 capsid proteins of bocavirus are encoded by a single RNA, VP1 and VP2 of PPV4 are encoded by two separate RNAs.

Detection of a novel porcine parvovirus, PPV4, in Chinese swine herds.

To determine whether the novel porcine parvovirus type 4 (PPV4) recently reported in America is prevalent in China, a set of specific primers was designed and used for molecular survey of PPV4 among the clinical samples collected from various provinces of China between 2006 and 2010. The results showed that PPV4 is present in Chinese swine herds at a rate of 2.09% (12/573) among the clinical samples examined and 0.

[Identification of differentially expressed proteins in the liver of Oncomelania snails induced by Eomecon chinanthe sanguinarine].

Objective: To identify the differentially expressed proteins in the liver of Oncomelania snails induced by Eomecon chinanthe sanguinarine.

Methods: Sanguinarine was extracted and purified from the dry powder of Eomecon chinanthe. Oncomelania snails were immersed in 5 mg/L sanguinarine (50 Oncomelania snails per 500 ml) or pure water for 36 h (25°C) and the livers were isolated from live snails.

2-, 3-, and 4-(1-Oxo-1H-2,3-dihydroisoindol-2-yl)benzoic acids and their corresponding organotin carboxylates: synthesis, characterization, fluorescent, and biological activities.

Three novel organotin complexes with general formula Sn(OH)(bz)(2)L (bz = benzyl, HL = 2-, 3-, or 4-(1-oxo-1H-2,3-dihydroisoindol-2-yl)benzoic acid) and one of their ligands were prepared and characterized. In vitro antifungal and antibacterial activities of these complexes and ligands were investigated with the representative strains of Candida albicans, Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa. Their fluorescence properties have also been discussed.

[Screening and identification of differentially expressed proteins between adult female and male worms of Schistosoma japonicum].

Objective: To screen and identify differentially expressed proteins between adult female and male worms of Schistosoma japonicum(S.japonicum).

Methods: Two rabbits infected with the cercaria were perfused with saline in carotid, and approximately two hundred adult female and two hundred male worms of S.

Construction of infectious cDNA clones of PRRSV: separation of coding regions for nonstructural and structural proteins.

Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), the causative agent of the ongoing "porcine high fever syndrome" in China, is capable of genetic and antigenic mutations at high frequency. How to design vaccine rationally to keep up with the ever-changing prevalent PRRSV variant is of great interest. We developed an infectious cDNA clone of an attenuated strain of Type II PRRSV, and further manipulated the infectious cDNA clone by inserting polylinker between ORF1 and ORF2, encoding for nonstructural-or structural-protein, respectively.

[Identification of porcine reproductive and respiratory syndrome virus regulation sequence in 3'-untranslated region].

Based on our established infectious clone of PRRSV, designated as pCBC2, a series of mutagenesis of 3'-untranslated region (3'-UTR) at primary structure and secondary structure level were constructed. Then the full length mutant clones were transfected into MARC-145 cells, from which the influences of the discrete 3'-UTR mutation on PRRSV replication and transcription were analyzed. The properties of the rescued mutant viruses were then further characterized by Northern Blot and plaque morphology analysis.

[Expression and purification of Toxoplasma gondii GRA4 gene in prokaryotic system].

Objective: To construct a prokaryotic expression system containing the dense granule protein 4 (GRA4) of Toxoplasma gondii, purify the expressed protein and detect its immunogenicity.

Methods: The specific fragment of GRA4 gene was amplified by PCR. After subcloning the prokaryotic expression recombinant pET-GRA4, the expressed product was purified with His Bind affinity chromatography and analyzed by Western blot.

[Expression of truncated Toxoplasma gondii GRA8 in the prokaryotic expression plasmids].

Objective: To construct the prokaryotic recombinant expression plasmids of Toxoplasma gondii GRA8 and analyze their expression in E. coli containing the prokaryotic recombinant plasmids.

Methods: The full gene and its truncated fragment of GRA8 were amplified by PCR from T.

[Cloning and characterization of new genes of Schistosoma japonicum].

Objective: To clone and characterize new genes of Schistosoma japonicum, Sj, and to provide efficient vaccine candidates.

Methods: Sj adult cDNA library was screened with rabbit sera raised against male worm soluble antigen. The inserted cDNA fragments from the positively selected clones were amplified with PCR and further sequenced, as well as characterized through internet NCBI GenBank software.

[Study on the subcloning, expression and immunoprotection with Schistosoma japonicum CAI gene].

Objective: To subclone and express the new gene of Schistosoma japonicum (Sj) CAI and evaluate the immunoprotective effect of the recombinant molecule.

Methods: The cDNA of SjCAI gene was subcloned into expression vector pGEX-5X-3 to form recombinants which were then used to transform to E. coli strain ER 2566.

[Immunoscreening of Schistosoma japonicum adult worm cDNA library with sera vaccinated with cercaria antigen and analysis of novel genes].

Objective: To explore antigens possessing common immunogenicity with Schistosoma japonicum (Sj) cercariae antigens, and to find out new candidate antigens for schistosomiasis diagnosis and vaccine.

Methods: Sj adult cDNA library was screened using sera from rabbits vaccinated with Sj cercariae antigen, the inserts of positive clones were amplified by PCR, all positive clones were sequenced and the data were analysed using Nucleotide BLAST software of NCBI and Expert Protein Analysis System of Swiss Institute of Bioinformatics.

Results: Thirteen positive clones were obtained after three rounds of immunoscreening, and all amplified by PCR.

[Study on diagnosis of schistosomiasis by ELISA using periodate-treated soluble egg antigen].

Objective: To improve SEA-ELISA, an immunodiagnostic assay for schistosomiasis.

Methods: Soluble egg antigen (SEA) of Schistosoma japonicum was treated with sodium periodate (SP) in order to oxidate its glycosylated epitopes. ELISA using the treated SEA was then performed to detect specific antibodies to SEA in the sera of schistosomiasis patients.

[Studies on fractionation and protective immunity of the membrane antigen from hepato-portal juveniles of Schistosoma japonicum].

Objective: To explore the immunological characteristics of the membrane antigen from hepato-portal juveniles of Schistosoma japonicum and its protective immunity against S. japonicum (Sj) in mice.

Methods: Sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) and enzyme-linked immune electro-transfer blot(EITB) methods were used to recognize the membrane antigens from hepato-portal schistosomula (SjHmAg) by infected rabbit sera (IRS) and normal rabbit sera (NRS).

Molecular cloning, expression and vaccination of a novel gene Sj-MA of Schistosoma japonicum.

In order to obtain new gene and to develop the new vaccine candidate of immunoprotection against Schistosoma japonicum (Sj), Sj adult worm cDNA library was screened with anti-sera to soluble male adult worm antigen and resulted in the discovery of a novel gene designated as Sj-MA. Sequence analysis showed that Sj-MA as a complete cDNA contains one open reading frame. It was deduced to contain 249 amino acid residues and encode a 28.

[Fractionation and identification of Schistosoma japonicum adult worm 67kD protein].

Objective: To isolate the specific protein bands which react to sera from rabbits infected with Schistosoma japonicum (Sj) and those immunized with Sj adult worm antigens (AWA) for immunodiagnosis of schistosomiasis.

Methods: AWA was analysed using SDS-PAGE and immunoblotting, and 67 kD protein was fractionated by electrophoretic chromatography.

Results: 67 kD protein which reacted to rabbit sera infected with Sj and immunized with AWA was obtained after the fractionation of AWA through electrophoretic chromatography.