In order to investigate the apoptosis triggered by porcine circovirus type 2 (PCV2) in lymphocytes and the underlying mechanism, the levels of apoptosis and the expression levels of miRNA were examined by flow cytometry, Western blotting and real-time PCR (qPCR). The mimics or inhibitors of miR-125a-5p, an apoptosis-related miRNA, were transfected into PK-15 cells, and the apoptosis rate was examined upon overexpression or inhibition of mir-125a-5p. The target gene of mir-125a-5p was predicted by bioinformatics method, and the regulation of mir-125a-5p on the target gene was analyzed by luciferase reporter assay. The expressions of Bcl-2, Bax, cytochrome C and caspase-3 were detected by Western blotting. The results showed that exosomes secreted by PK-15 cells infected with PCV2 significantly increased the lymphocyte apoptosis rate, which was dose-dependent in certain concentration range. The expression of miR-125a-5p was dramatically increased. The apoptosis rate was increased significantly in the cells transfected with miR-125a-5p. It was predicted that there were binding sites of miR-125a-5p at Bcl-2 3'UTR by TargetScan. The luciferase activity of wild-type pmir-Bcl-2 3'UTR was inhibited significantly by miR-125a-5p mimics, but that of mutant pmir-Bcl-2 3'UTR was not changed. By Western blotting, Bcl-2 was reduced significantly, while Bax, cytochrome C and caspase-3 increased significantly, and the ratio of Bcl-2/Bax was significantly decreased. These results showed that PCV2 up-regulated the expression of miR-125a-5p through exosomes, then inhibited the expression of Bcl-2 at both mRNA and protein level, activated mitochondrial apoptosis pathway and induced apoptosis in lymphocytes.
3' Untranslated Regions ; Animals ; Apoptosis/genetics* ; Caspase 3/genetics* ; Cell Proliferation/genetics* ; Circovirus/genetics* ; Cytochromes c/genetics* ; Luciferases/genetics* ; Lymphocytes/metabolism* ; MicroRNAs/metabolism* ; Swine ; bcl-2-Associated X Protein/genetics*

Objective:To investigate the level of rumination and its influential factors among Chinese psychiatry nurses who go through work place violence.Methods:In this study, 150 psychiatry nurses were recruited from Affiliated Brain Hospital of Guangzhou Medical University (specialize in psychiatry), via the combination of convenient sampling and snowball sampling. Chinese Event-related Rumination Inventory (C-ERRI) was applied in the survey.Results:The total score of C-ERRI was (22.11±9.62) points, and the scores of intrusive rumination subscale and deliberate rumination subscale were (12.99±5.58) and (9.12±6.01) points, respectively. Multiple Linear Regression Analysis show that assault frequency in recent year ( B=-3.195, P<0.01) and whether got injury in the recent assault ( B=8.591, P<0.01) were predictors of deliberate rumination, which account for 26.8% variance of the equation. Gender (male) ( B=-2.415, P<0.01), Injury frequency in recent year ( B=2.864, P<0.01) and whether got injury in the recent assault ( B=8.949, P<0.01) were predictors of intrusive rumination. They account for 36.0% variance of the equation. Conclusion:In this study, the level of rumination among Chinese psychiatry nurses was low. Their rumination style was deliberate rumination.

Objective:To explore problems and self-experience of the newly graduated recruited nurses in the process of work and to provide a basis for formulating solutions to allow newly graduated recruited nurses to adapt to clinical work as soon as possible.Methods:The phenomenological method of qualitative study was used to interview 25 newly recruited nurses in a ClassⅢ Grade A hospital in Guangzhou from 2018 to 2019.Results:By sorting out, refining and analyzing the interview data, three concepts related to professional confusion, career planning limitation and emphasis on professional acquisition were extracted.Conclusions:Professional confusion is common in newly recruited nurses in department of psychiatry. Medical staff should pay attention to the psychological status and self-experience of newly recruited nurses in department of psychiatry, provide necessary help and support and reduce the negative experience of nurses so as to make them better adapt to clinical work.

Recombinant PRRSV △2ORF5 gene was constructed using DNA shuffling from four genetically different strains of PRRSV to study its heterologous cross-neutralizing ability. The △2ORF5 mutant gene was cloned into the vector pET-32a and transferred into E. coli BL21. SDS-PAGE confirmed that the molecular weight of the recombinant △2ORF5 was about 42 kDa, consistent with the predicted result. Then the purified recombinant protein was injected into BALB/c mouse to obtain polyclonal antibody. Western blotting analysis with mouse-anti-△2ORF5 polyclonal serum indicated that the parental virus recombinant GP5 protein reacted with the specific antibodies. Four parental viruses could be inhibited by the anti-△2ORF5 polyclonal antibody and the inhibition rates were higher than 53%. This work has laid a foundation for further development vaccine for PRRSV.

Objective To evaluate the diagnostic efficiency of deep fungal infection by detecting the serum galactomannan ( GM) and bronchoalveolar lavage fluid ( BALF) GM, serum G test and fungal culture of BALF in patients with suspected invasive fungal infection ( IFI) in lungs.Methods A retrospective analysis was performed of the results of serum /BALF GM test ,serum G test and BALF culture from 148 patients with suspected pulmonary IFI .The indexes involved sensitivity , specificity , positive predictive value , negative predictive value , as well as diagnostic capacity for deep fungal infection with separated or combined tests .Results Among the 148 cases, 48 cases were clinically diagnosed with IFI and the rest were excluded.Among the 48 IFI cases, 3 cases were positive in serum GM test , 25 cases were positive in BALF GM test , 31 cases were positive in G test and 30 cases were positive in fungal culture .The combined detection showed a sensitivity of 91.6%,specificity of 70.0%, positive predictive value of 59.5% and negative predictive value of 94.6%.Conclusion The combination of GM/G tests and fungal culture can significantly improve the clinical diagnostic efficiency of pulmonary IFI .

Objective To investigate the laboratory diagnosis of filamentous fungi which are rarely seen in clinical practice.Methods Five strains of fungi were isolated from clinical samples and initially identified by the morphological method.Total DNA of fungi was extracted and amplified by the PCR method using universal primers of ITS2-ITS4 gene, respectively.The PCR products were sequenced and the obtained sequences were then analyzed by the blastn program incorporated in NCBI.Results The five strains of fungi were diagnosed as Scedosporium apiospermum,Schizophyllum commune,Scopulariopsis brevicaulis,Rhizopus stolonifer,and Fusarium solani.Conclusion The laboratory diagnosis of filamentous fungi which rarely occur in clinical practice should integrate various methods,including morphological, microbiological,and molecular biological methods.

Objective To investigate the flora distribution and drug resistance status in the Affiliated Hospital of Academy of Military Medical Sciences so as to provide experimental data for clinical doctors to use antibiotics more efficiently.Methods The clinical data of pathogenic bacterial infections over nearly one year in our hospital were retro -spectively analyzed .Results There were 3815 strains of pathogenic bacteria isolated from the sample .The percentage of Gram-positive strains was 36.4%while that of Gram-negative bacteria was 63.6%.The most common bacteria were Esche-richia coli, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and Staphylococcus epidermidis.In terms of drug tolerance , Enterobacteriaceae remained highily sensitive to carbapenems .The total resistance rate was 2%-5%.The resistance rate of A.baumannii to meropenem and imipenem was 60%.There were still a few pan-drug resistant strains among K.pneumoniae,A.baumannii and P.aeruginosa,but there were no drug resistant strains to vancomycin , tige-cycline and linezolid in Staphylococcus.The resistance rate of Enterococcus faecium to vancomycin was 9%.The bacteria were distributed predominantly in ICU ,Department of Hematology and Department of Oncology .The samples were mainly composed of phlegm specimens .Conclusion The high distribution in the three departments mentioned above is largely re-lated to the diseases being treated .The specimens from the lower respiratory tract show more types of bacteria that are mostly drug-resistant, and the isolating rate of vancomycin resistant Enterococcus and carbapenems resistant K.pneumoniat is com-paratively high .

Many researchers employed mammalian expression system to artificially express cannabinoid receptors, but immunoblot data that directly prove efficient protein expression can hardly be seen in related research reports. In present study, we demonstrated cannabinoid receptor protein was not able to be properly expressed with routine mammalian expression system. This inefficient expression was rescued by endowing an exogenous signal peptide ahead of cannabinoid receptor peptide. In addition, the artificially synthesized cannabinoid receptor was found to aggregate under routine sample denaturing temperatures (i.e., [Symbol: see text]95°C), forming a large molecular weight band when analyzed by immuno-blotting. Only denaturing temperatures [Symbol: see text]75°C yielded a clear band at the predicted molecular weight. Collectively, we showed that efficient mammalian expression of cannabinoid receptors need a signal peptide sequence, and described the requirement for a low sample denaturing temperature in immuno-blot analysis. These findings provide very useful information for efficient mammalian expression and immuno-blotting of membrane receptors.

Many researchers employed mammalian expression system to artificially express cannabinoid receptors,but immunoblot data that directly prove efficient protein expression can hardly be seen in related research reports.In present study,we demonstrated cannabinoid receptor protein was not able to be properly expressed with routine mammalian expression system.This inefficient expression was rescued by endowing an exogenous signal peptide ahead of cannabinoid receptor peptide.In addition,the artificially synthesized cannabinoid receptor was found to aggregate under routine sample denaturing temperatures (i.e.,≥95℃),forming a large molecular weight band when analyzed by immuno-blotting.Only denaturing temperatures ≤75℃ yielded a clear band at the predicted molecular weight.Collectively,we showed that efficient mammalian expression of cannabinoid receptors need a signal peptide sequence,and described the requirement for a low sample denaturing temperature in immuno-blot analysis.These findings provide very useful information for efficient mammalian expression and immuno-blotting of mcmbrane receptors.

We isolated a novel Enterobacteria phage IME08 from hospital sewage, then confirmed it was a double-stranded DNA phage by digesting its genetic material with DNase I, RNase A and several restriction endonucleases respectively. BLAST results of random fragments generated by a random PCR cloning method revealed that it belonged to T4-like virus. We subsequently determined the host recognizing genes (g37 and g38) sequence with a PCR-based "genome jumping" protocol based on highly conserved region at 5' terminus of g37 from four other T4-like Bacteriophages (T4, JS98, T2 and K3). These molecular biological methods enabled us to readily characterize the bacteriophage and efficiently determine the sequence of the genes of interest based on very limited conserved sequence information.
Bacteriophage T4 ; genetics ; isolation & purification ; Cloning, Molecular ; DNA, Viral ; genetics ; Escherichia coli ; genetics ; virology ; Genome, Viral ; genetics ; Host Specificity ; genetics ; Polymerase Chain Reaction ; methods