1.Unilateral pedicle screw fixation combined with lumbar interbody fusion for the treatment of lumbar degenerative diseases
Zhanyong WU ; Yongcheng HU ; Yundong WEI ; Hualong WU ; Xiangping PENG ; Jianjun KONG ; Laibao YU ; Shaofeng WANG ; Shuangtao CHEN ; Jianguo SUN
Chinese Journal of Orthopaedics 2010;30(11):1109-1115
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.
2.Diagonsis establishment of fluorescen quantitative PCR assay for pseudorabies wild-type virus and vaccine virus.
Li ZHAO ; Baoan CUI ; Hongying CHEN ; Zhanyong WEI ; Lanlan ZHENG ; Xiaoli LÜ ; Yanyan JIA ; Xuyong ZHAO
Chinese Journal of Biotechnology 2008;24(7):1149-1154
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
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Fluorescent Dyes
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Herpesvirus 1, Suid
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genetics
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isolation & purification
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Polymerase Chain Reaction
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methods
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Pseudorabies
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diagnosis
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prevention & control
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virology
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Pseudorabies Vaccines
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immunology
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isolation & purification
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Swine
3.Gene cloning, expression and activity detection of porcine interleukin-18 mature protein in Escherichia coli.
Lanlan ZHENG ; Yunfei JIA ; Baoan CUI ; Hongying CHEN ; Zhanyong WEI ; Ruiliang CHEN
Chinese Journal of Biotechnology 2008;24(2):214-219
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
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Animals
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Base Sequence
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Cloning, Molecular
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Escherichia coli
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genetics
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metabolism
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Interleukin-18
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biosynthesis
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genetics
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Molecular Sequence Data
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Recombinant Fusion Proteins
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biosynthesis
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genetics
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immunology
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Swine
4.Porcine parvovirus nonstructural protein NS1 activates NF-κB and it involves TLR2 signaling pathway
Xiaohui JIN ; Yixin YUAN ; Chi ZHANG ; Yong ZHOU ; Yue SONG ; Zhanyong WEI ; Gaiping ZHANG
Journal of Veterinary Science 2020;21(3):e50-
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.
5.Antiviral effects of Bovine antimicrobial peptide against TGEV in vivo and in vitro
Xiuli LIANG ; Xiaojun ZHANG ; Kaiqi LIAN ; Xiuhua TIAN ; Mingliang ZHANG ; Shiqiong WANG ; Cheng CHEN ; Cunxi NIE ; Yun PAN ; Fangfang HAN ; Zhanyong WEI ; Wenju ZHANG
Journal of Veterinary Science 2020;21(5):e80-
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.