Analysis of Detection of Mixed Semen Stains with Different Immunological Test Methods and Case Applications / 法医学杂志

Objective To perform the separation and confirmation of mixed semen stains with immunological test method, and find a more effective method for the detection of mixed semen stains. Methods The semens of three volunteers were mixed. The mixed semen stains were processed and tested with prostate-specific antigen （PSA） colloidal gold immunoassay strip method, immunomagnetic beads method and laser capture microdissection, respectively. Statistics of the results of STR were gathered and compared with those of a single semen stain. Results After PSA colloidal gold immunoassay strip method testing, the samples showed a purplish red line in the test area and the control area. The results obtained with the immunomagnetic beads method showed a more complete and effective short tandem repeat （STR） sequence. The mixed semen stains were processed with laser capture microdissection and low volume amplified. The results were summarized and superimposed to obtain a complete single typing, which matched the single semen stain typing, with a typing success rate of 84.00%. Single suspect Y-STR typing was obtained with the application of the method above in actual cases, which provided evidence basis for rapid solving of the case. Conclusion The combination of PSA colloidal gold immunoassay strip method, immunomagnetic beads method and laser capture microdissection can be used to separate and confirm the mixed semen stains.

Herpesviral infection and Toll-like receptor 2

In the last decade, substantial progress has been made in understanding the molecular mechanisms involved in the initial host responses to viral infections. Herpesviral infections can provoke an inflammatory cytokine response, however, the innate pathogen-sensing mechanisms that transduce the signal for this response are poorly understood. In recent years, it has become increasingly evident that the Toll-like receptors (TLRs), which are germline-encoded pattern recognition receptors (PRRs), function as potent sensors for infection. TLRs can induce the activation of the innate immunity by recruiting specific intracellular adaptor proteins to initiate signaling pathways, which then culminating in activation of the nuclear factor kappa B (NF-κB) and interferon-regulatory factors (IRFs) that control the transcription of genes encoding type I interferon (IFN I) and other inflammatory cytokines. Furthermore, activation of innate immunity is critical for mounting adaptive immune responses. In parallel, common mechanisms used by viruses to counteract TLR-mediated responses or to actively subvert these pathways that block recognition and signaling through TLRs for their own benefit are emerging. Recent findings have demonstrated that TLR2 plays a crucial role in initiating the inflammatory process, and surprisingly that the response TLR2 triggers might be overzealous in its attempt to counter the attack by the virus. In this review, we summarize and discuss the recent advances about the specific role of TLR2 in triggering inflammatory responses in herpesvirus infection and the consequences of the alarms raised in the host that they are assigned to protect.

Cloning, prokaryotic expression and subcellular localization in the infected host cells of the duck plague virus DPV UL35 gene / 病毒学报

Based on the duck plague virus (DPV) UL35 gene sequence that our laboratory obtained (GenBank accession number EF643558), a pair of primers was designed using Oligo6.0 and primer5.0, then the UL35 gene was amplified from DPV CHv strain genomic DNA and cloned into the pMD18-T to construct a clone plasmid pMD18-T-UL35. After identification of the pMD18-T-UL35 by PCR amplification and restriction digestion, the fragment of the UL35 gene was subcloned into the prokaryotic expression vector pET-32a(+). The resultant recombinant plasmid pET-32a(+)-UL35 was then transformed into E. coli BL21 (DE3) strain and optimally-expressed under the induction of 1.0 mmol/L IPTG at 34 degrees C for 5 hours. SDS-PAGE analysis showed the recombinant protein (VP26) had a molecular weight of about 33KDa and accounted for 32.3% of total bacterial protein by gel scanning. The protein was then purified by Ni(2+)-affinity chromatography and used to immunize rabbit for producing the VP26 anti-serum and its antibody titer was up to 1:32 detected by agar diffusion reaction. After the IgG of the polyclonal antibodies was purified by High-Q anion-exchange chromatography, Western blot analysis indicated that the IgG had specific reaction with the VP26. Moreover, the subcellular localization detection was observed using immunofluorescence technique. The results showed that the specific fluorescences appeared relatively few in nucleus in 2 to 8 hours and increased gradually in 12 to 36 hours and eventually reached to the maximum, which aggregated in the spot region of the nucleus after the duck embryo fibroblast (DEF) were infected by DPV. However, there were only a small amount of specific fluorescences in the cytoplasm in 12 hours and increased with the extension of infection time in 24 to 48 hours. The specific fluorescences finally reached to the maximum in the cytoplasm in 72 hours. The results provided significant data for furthering the study on the function of DPV UL35 gene.