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Objective:To evaluate the effect of vitamin K 2 on traumatic brain injury (TBI) and the relationship with nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3 (NLRP3) inflammasomes in rats. Methods:Thirty-six SPF healthy male Sprague-Dawley rats, weighing 280-300 g, were divided into 3 groups ( n=12 each) by a random number table method: sham operation group (group Sham), group TBI and TBI plus vitamin K 2 group (group TBI+ VK 2). The TBI model was developed using modified Feeney′s method.In TBI+ VK 2 group, vitamin K 2 400 mg/kg (dissolved in dimethyl sulfoxide) was intraperitoneally injected at 30 min after developing TBI model.The equal volume of dimethyl sulfoxide was intraperitoneally injected in group Sham and group TBI.The modified neurological severity score (mNSS) was measured and open field tests were performed at 24 h after development of TBI.The rats were sacrificed after the end of behavioral testing, and brains were obtained for measurement of brain water content (by wet-dry weight method), percentage of brain injury volume (by TTC assay), contents of interleukin-1β (IL-1β), IL-18 and caspase-1 in cortex on the injured side (by enzyme-linked immunosorbent assay) and expression of NLRP3, caspase-1 and IL-18 in cortex on the injured side (by Western blot). Results:Compared with group Sham, the mNSS score was significantly increased, the total distance travelled was reduced, the time spent in the central zone was shortened, the brain water content and percentage of brain injury volume were increased, the contents of IL-1β, IL-18 and caspase-1 in cortex on the injured side were increased, and the expression of NLRP3, caspase-1 and IL-18 was up-regulated in group TBI ( P<0.05 or 0.01). Compared with group TBI, the mNSS score was significantly decreased, the total distance travelled was increased, the time spent in the central zone was prolonged, the brain water content and percentage of brain injury volume were decreased, the contents of IL-1β, IL-18 and caspase-1 in cortex on the injured side were decreased, and the expression of NLRP3, caspase-1 and IL-18 was down-regulated in group TBI+ VK 2 ( P<0.05 or 0.01). Conclusions:Vitamin K 2 can reduce TBI, and the mechanism may be related to inhibition of the activation of NLRP3 inflammasomes in rats.
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Objective To evaluate the safety and drug adherence of tocilizumab(TCZ)in patients with moderate to severe rheumatoid arthritis(RA)in routine clinical practice. Methods This 24 week single center observational study recruited patients with moderate to severe RA. Therapy adherence rate was calculated by actual dosing/expected dosing×100%. Efficacy end points included physician global assessment of disease activity(PGA),patient global assessment of disease activity(PtGA),28-joint disease activity score(DAS28)and so on. Safety was evaluated by recorded adverse events (AEs). Results Sixty patients were enrolled with a mean (SD) treatment adherence of (67±27)%. PGA, PtGA, pain assessment (VAS), TJC and SJC all decreased during this study. At the 12th week, 25%(6/24) and 29%(7/24) of the patients achieved DAS28 remission and EULAR good response,respectively.Eighteen AEs were recorded,of which only 2 were severe AEs(SAEs)and neither was related to TCZ. Conclusion TCZ is a highly safe treatment for decreasing disease activity in patients with moderate to severe RA in China.However,drug adherence still need to be improved.
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This study was aimed to establish an HPLC method to determine three ginsenosides in Shengmai ultra-micro powder. The kromasil C18 (250 mm í 4.6 mm, 5 μm) was used as analytical column. The mobile phase was composed of acetonitrile (A) and water (B) with gradient elution (0~35 min, 19% A; 35~55 min, 19%~29% A; 55~70 min, 29% A; 70~100 min, 29%~40% A) at a flow rate of 1 mL·min-1. The detection wavelength was 203 nm and the column temperature was 30℃. The injection volume was 10 μL. The results showed that the linear ranges of ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1 were 0.083~0.834 μg, 0.086~0.863 μg, 0.091~0.911 μg, respec-tively. The average recovery rates (n = 6) were 100.7%, 100.5%, 100.5%, respectively. It was concluded that this method was quick, sensitive, repeatable and suitable to determine contents of ginsenoside Rg1, ginsenoside Re and ginsenoside Rb1 in Shengmai ultra-micro powder.
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The aim of the study is to establish a new method of quality evaluation and validate its feasibilities by the simultaneous quantitative assay of four lignanoids in Schisandra chinensis. A new quality evaluation method, quantitative analysis of multi-components by single marker (QAMS), was established and validated with Schisandra chinensis. Four main lignanoids, schisandrin, schisantherin A, deoxyschizandrin and gamma-schizandrin, were selected as analytes and schisandrin as internal reference substance to evaluate the quality. Their contents in 13 different batches of samples, collected from different bathes, were determined by both external standard method and QAMS. The method was evaluated by comparison of the quantitative results between external standard method and QAMS. No significant differences were found in the quantitative results of four lignanoids in 13 batches of S. chinensis determined by external standard method and QAMS. QAMS is feasible for determination of four lignanoids simultaneously when some authentic standard substances were unavailable, and the developed method can be used for quality control of S. chinensis.
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Objective To investigate the mRNA expression level of neurogranin on peripheral blood mononuclear cells(PBMCs)from patients with systemic lupus erythematosus(SLE).Methods Top 20 tags of SLE PBMCs SAGE library were searched from normal lymphocytes SAGE library including navie-T,Th1,Th2, CD8+T, NK and B cells,and their abundance was compared.The mRNA expression level of neuro-granin,a differential over-expressed tag,was detected in 35 cases of SLE and 15 normal controls by reversetranscription-polymerase chain reaction (RT-PCR).Results Neurogranin tag could only be detected in SLE PBMCs SAGE library,but was hardly found in normal lymphocyte SAGE library.However,either SLE pa-tients or normal controls showed a detectable mRNA level of neurogranin on PBMCs by RT-PCR.The mRNA level of neurogranin in active SLE patients was significantly increased than those in controls(P＜0.001).but only slightly increased in inactive SLE patients (P＞0.05).Conclusion Neurogranin,as a novel proapototic factor,is overexpressed on PBMCs of SLE patients.It may be involved in the regulation of abnormal immune responses in lupus.
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This study examined the gene expression patterns of peripheral blood mononuclear cells (PBMCs) in patients with systemic lupus erythematosus (SLE) by using serial analysis of gene expression (SAGE) technology. Following the construction of serial analysis of gene expression (SAGE) library of PBMCs collected from 3 cases of familial SLE patients, a large scale of tag sequencing was performed. The data extracted from sequencing files was analyzed with SAGE 2000 V 4.5 software. The top 30 expressed genes of SLE patients were uploaded to http://david.niaid.nih.gov/david/ease.htm and the functional classification of genes was obtained. The differences among those expressed gene were analyzed by Chi-square tests. The results showed that a total of 1286 unique SAGE tags were identified from 1814 individual SAGE tags. Among the 1286 unique tags, 86.8% had single copy, and only 0.2% tags had more than 20 copies. And 68.4% of the tags matched known expressed sequences, 41.1% of which matched more than one known expressed sequence. About 31.6% of the tags had no match and could represent potentially novel genes. Approximately one third of the top 30 genes were ribosomal protein, and the rest were genes related to metabolism or with unknown functions. Eight tags were found to express differentially in SAGE library of SLE patients. This study draws a profile of gene expression patterns of PBMCs in patients with SLE. Comparison of SAGE database from PBMCs between normal individuals and SLE patients will help us to better understand the pathogenesis of SLE.
Subject(s)
Expressed Sequence Tags , Gene Expression Profiling/methods , Leukocytes, Mononuclear/metabolism , Lupus Erythematosus, Systemic/blood , Lupus Erythematosus, Systemic/genetics , Sequence Tagged Sites , Transcription, GeneticABSTRACT
This study examined the gene expression patterns of peripheral blood mononuclear cells (PBMCs) in patients with systemic lupus erythematosus (SLE) by using serial analysis of gene expression (SAGE) technology. Following the construction of serial analysis of gene expression (SAGE) library of PBMCs collected from 3 cases of familial SLE patients, a large scale of tag sequencing was performed. The data extracted from sequencing files was analyzed with SAGE 2000 V 4.5 software.The top 30 expressed genes of SLE patients were uploaded to http://david.niaid.nih. gov/david/ease.htm and the functional classification of genes was obtained. The differences among those expressed gene were analyzed by Chi-square tests. The results showed that a total of 1286 unique SAGE tags were identified from 1814 individual SAGE tags. Among the 1286 unique tags, 86.8% had single copy, and only 0.2% tags had more than 20 copies. And 68.4% of the tags matched known expressed sequences, 41.1% of which matched more than one known expressed sequence. About 31.6% of the tags had no match and could represent potentially novel genes. Approximately one third of the top 30 genes were ribosomal protein, and the rest were genes related to metabolism or with unknown functions. Eight tags were found to express differentially in SAGE library of SLE patients. This study draws a profile of gene expression patterns of PBMCs in patients with SLE. Comparison of SAGE database from PBMCs between normal individuals and SLE patients will help us to better understand the pathogenesis of SLE.
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To develop a method for identification of differential gene expression between different cell populations, several convenient techniques of molecular biology, including subtractive hybridization, suppression PCR, T/A cloning and sequencing, were used to identify genes expressed differentially in CD45+ and CD45- cells isolated from U266 cell line of multiple myeloma. Our results showed that the levels of abundant genes scale down 20 times through subtractive hybridization. Plasmid DNA from CD45- cell clones was hybridized with forward or backward cDNA probes synthesized from CD45- and CD45- cells, respectively. A few of differentially expressed genes reconfirmed by RT-PCR were identified from 500 expressed clones of CD45+ cells. It is concluded that a strategy for gene expression identification developed from conventional molecular biological methods can be used in different laboratories.
Subject(s)
Humans , Cell Line, Tumor , DNA , Genetics , Gene Expression Profiling , Gene Expression Regulation, Neoplastic , Leukocyte Common Antigens , Genetics , Multiple Myeloma , Genetics , Pathology , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
Objective To evaluate the effect of double interventional thrombolysis for treatment of acute venous thrombosis in lower limb and discuss the problems of its clinical use. Methods 24 patients with acute venous occlusive disease associated with thrombus formation were treated by double interventional thrombolysis including 17 cases of femoral venous approach and 7 with femoral artery catheterization. The total dosage of urokinase was from 2 500 000 U to 5 000 000 U. Results All together 24 cases were undengone this proceduce; outcoming with complete thrombolysis in18(75%), partial and noneffective in 3(12.5%) and 3 (12.5%) respectively. The total effective rate was 87.5%. Conclusions Double interventional thrombolysis is highly effective to the patients with acute venous thrombosis.