Objective To explore the feasibility and efficiency of the treatment of lumbar degenerative diseases after transforaminal lumbar interbody fusion(TLIF)and posterolateral fusion(PLF)procedures in which unilateral pedicle screw fixation was used.Methods From December 2006 to August 2008,78 cases with the lumbar degenerative diseases who received lumbar posterolateral fusion were analyzed.There were 48 cases of which underwent TLIF and PLF procedures with unilateral pedicle screw fixation(unilateral group),including 25 males and 23 females with an average of 47.6 years;and 30 cases of which underwent TLIF and PLF procedures with bilateral pedicle screw fixation(bilateral group),including 21 males and 9 females with an average of 50.5 years.The clinical effects between the two groups were evaluated with Oswestry disability index and visual analogue score(VAS)index.The operation time,blood loss,fusion rates and intervertebral collapse rates were also compared.Results Oswestry disability index,low back pain VAS index and skelalgia VAS index in both groups showed statistical significance between preoperation and 3 months,or 3 months and 1 year postoperatively.There was no difference in score improvement between the two groups.There were difference in operation time,blood loss and cost of hospitalization between unilateral and bilateral group.The former was lower.There was no difference in postoperative length of stay between the two groups.The fusion rate of unilateral group and bilateral group were 91.7%(44/48)and 93.3%(28/30),respectively.Conclusion Auto graft combined with unilateral pedicle screw fixation provids better spinal instant stability.TLIF and PLF with unilateral pedicle screw fixation was a satisfactory method in treating degenerative disease of lumbar vertebrae.

Porcine interleukin-18 mature protein gene was amplified from porcine spleen cells by RT-PCR. PCR product was cloned into the T vector pGEM-T for sequencing. The nucleotide sequence of this gene was 474 bp. Then, it was subcloned into the prokaryotic expressing plasmid vector pGEX6P-1 and transformed into host E. coli strain BL21 for expression. The expression of pIL-18 mature protein gene was identified by SDS-PAGE .The expression product was fusion protein with molecular weight of 45 kD and the percentage of expression protein in E. coil protein was 28%. The protein was purified by washing of inclusion bodies and the activity was measured by methyl thiazolyl tetrazolium (MTT).
Amino Acid Sequence ; Animals ; Base Sequence ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Interleukin-18 ; biosynthesis ; genetics ; Molecular Sequence Data ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Swine

We designed two pairs of primers and their corresponding TaqMan probes according to gH, gE gene of PRV. By optimizing the probe's concentration, Mg2+ concentration, primers concentration and sample DNA extraction, real-time fluorescent quantitative PCR (FQ-PCR) which can quickly identity field virus and vaccine virus of PRV was established. According to our results, the dynamic range of the FQ-PCR assay is between 10 x 10(1) copies/microL and 10 x l0(8) copies/microL, and the detection limit of FQ-PCR is 1.0 x 10(1) copies/microL, which is 100 fold higher than that of conventional PCR. We detected 60 doubtful tissue samples using the FQ-PCR assay, serum neutralization and conventional PCR. In conclusion, the FQ-PCR method is rapid, sensitive, specific and accurate, and can be used to detect field strains of PRV rapidly. The closed-tube format of the assay minimized the risk of contamination of subsequent reaction and the assay can be performed in 2 h or less. Development of real-time quantitative PCR provides the basis for the early and rapid detection and analyzing quantitatively the infectious degree of PRV.
Animals ; Fluorescent Dyes ; Herpesvirus 1, Suid ; genetics ; isolation & purification ; Polymerase Chain Reaction ; methods ; Pseudorabies ; diagnosis ; prevention & control ; virology ; Pseudorabies Vaccines ; immunology ; isolation & purification ; Swine

Background: Porcine parvovirus (PPV) is a single-stranded DNA virus that causes porcine reproductive failure. It is of critical importance to study PPV pathogenesis for the prevention and control of the disease. NS1, a PPV non-structural protein, is participated in viral DNA replication, transcriptional regulation, and cytotoxicity. Our previous research showed that PPV can activate nuclear factor kappa B (NF-κB) signaling pathway and then up-regulate the expression of interleukin (IL)-6. Objectives: Herein, the purpose of this study is to determine whether the non-structural protein NS1 of PPV also has the same function. Methods: Real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR), enzyme-linked immunosorbent assay, western blot, immunofluorescence assay and small interfering RNA (siRNA) were used. Results: Our findings demonstrated that PPV NS1 protein can up-regulate the expression levels of IL-6 and tumor necrosis factor-alpha in a dose-dependent manner. Moreover, PPV NS1 protein was found to induce the phosphorylation of IκBα, then leading to the phosphorylation and nuclear translocation of NF-κB. In addition, the NS1 protein activated the upstream pathways of NF-κB. Meanwhile, TLR2-siRNA assay showed TLR2 plays an important role in the activation of NF-κB signaling pathway induced by PPV-NS1. Conclusions These findings indicated that PPV NS1 protein induced the up-regulated of IL-6 expression through activating the TLR2 and NF-κB signaling pathways. In conclusion, these findings provide a new avenue to study the innate immune mechanism of PPV infection.

When eukaryotic cells are undergoing stimulations like oxidative stress,heat shock,or vi-ral infection,cellular stress response will quickly initiate to stop translation,leading to the produc-tion of a large number of stress granules(SGs).SGs,as dynamic membraneless protein-RNA ag-gregates,play an important role in maintaining cellular homeostasis and regulating host gene ex-pression.Upon viral invasion,the host halts translation to form SGs,thereby arresting the transla-tion of viral proteins within host cells.This process recruits innate immune-related host factors,limiting the virus from exploiting host translation machinery to complete its life cycle.It activates the innate immune pathway.However,some viruses have evolved multiple mechanisms to counter SGs,in order to provide a better intracellular environment for viral replication.Here,we review the generation of SGs,antiviral mechanism of SGs,and the strategies for escaping SGs during virus e-volution,in order to provide a new perspective for the development of antiviral drugs.

Background: In suckling piglets, transmissible gastroenteritis virus (TGEV) causes lethal diarrhea accompanied by high infection and mortality rates, leading to considerable economic losses. This study explored methods of preventing or inhibiting their production.Bovine antimicrobial peptide-13 (APB-13) has antibacterial, antiviral, and immune functions. Objectives: This study analyzed the efficacy of APB-13 against TGEV through in vivo and in vitro experiments. Methods: The effects of APB-13 toxicity and virus inhibition rate on swine testicular (ST) cells were detected using 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT). The impact of APB-13 on virus replication was examined through the 50% tissue culture infective dose (TCID50 ). The mRNA and protein levels were investigated by real-time quantitative polymerase chain reaction and western blot (WB). Tissue sections were used to detect intestinal morphological development. Results: The safe and effective concentration range of APB-13 on ST cells ranged from 0 to 62.5 µg/mL, and the highest viral inhibitory rate of APB-13 was 74.1%. The log10 TCID50 of 62.5 µg/mL APB-13 was 3.63 lower than that of the virus control. The mRNA and protein expression at 62.5 µg/mL APB-13 was significantly lower than that of the virus control at 24 hpi. Piglets in the APB-13 group showed significantly lower viral shedding than that in the virus control group, and the pathological tissue sections of the jejunum morphology revealed significant differences between the groups. Conclusions APB-13 exhibited good antiviral effects on TGEV invivo and in vitro.

Coronavirus infection can cause serious respiratory and digestive system diseases in hu-mans and animals.In recent years,the frequent outbreaks and newly outbreaks of coronavirus in-fection have threatened global public health and the development of livestock and poultry.Howev-er,the development of anti-coronavirus drugs and vaccines was restricted due to the insufficient understanding of the mechanisms of coronavirus pathogenesis and cross species transmission.Re-verse genetic manipulation technology is a powerful tool in virological research,which can be used for the study of pathogenesis mechanisms,replication mechanisms and function analysis of protein.It is also be used in the development of attenuated or gene labeled vaccines,and antiviral drugs.Due to the large genome and complex structure of the coronavirus,the reverse genetic manipulation technology of coronavirus has been lagging for a long time.With the continuous updating of molec-ular biology methods,various new construction strategies have emerged.This article focuses on the construction strategy of the reverse genetic operating system for coronavirus,as well as its applica-tion in virus transmission and pathogenic mechanisms,and development of vaccines,which will provide favorable tools for the prevention and control of the coronavirus infection.