The detection of HLA-B27 gene and gene typing of ankylosing spondylitis patients by fluorescence PCR / 国际检验医学杂志

Objective To investigate the clinical application value of fluorescence polymerase chain reaction (PCR) .in the human leucocyte antigen-B27(HLA-B27) gene and gene typing detection of ankylosing spondy-litis (AS) patients .Methods A total of 43 clinical blood samples of AS and 56 samples of healthy controls were collected in Shenzhen Futian hospital for rheumatic diseases from January 2014 to March 2015 .HLA-B27 gene was detected by flow cytometry .HLA-B27 gene and gene typing was also detected by the fluorescence PCR method .Results Among 43 samples ,40 samples were HLA-B27 positive(93 .02%) by flow cytometry while 39 samples were HLA-B27 positive (90 .70%) by fluorescence PCR .The total coincidence rate was 97 .50% .Among 39 positive samples ,32 samples were HLA-B2704 positive (82 .05%) and 7 samples were HLA-B2705 positive (17 .95%) .Conclusion The fluorescence PCR is an accurate method to detect HLA-B27 gene and presents high consistency with flow cytometry .It can also detect the HLA-B27 gene typing .It may have great clinical application value and prospects .

miR-155 facilitates the differentiation of Th17 cells by inhibiting the gene expression of Ets-1 / 中华风湿病学杂志

Objective To elucidate the function way of micro RNA(miR)-155 in the differentiation of Th17 cells.Methods CD4+T cells were separated from mice spleens using MACS CD4+T cells separatinge kit and cultured with interleukins [interleukin (IL)-2, IL-23 and IL-6] which could induce CD4+ T cells differentiate into Th17 cells.IL-17 was detected by flow cytometry and enzyme linked immunosorbent assay (ELISA) after transfected with miR-155 mimics or inhibitor lentiviral vectors.The expression levels of miR-155, IL-17A mRNA and Ets-1 mRNA were detected using fluorescent quantitation real-time quantitative polymerase chain reaction (RT-PCR).The si-Ets and miR-155 co-function for Th17 differentiation was analyzed.Data analysis was perfoemed using one-way analysis of variance (ANOVA) test and Dunnett test for pair-wise comparison and t test.P＜0.05 was considered to be statistically significant.Results The CD4+T cells were divided into four groups (the untreated control untreat group, the treatment control treat group, the miR-155 mimnics group and miR-155 inhibitor group).IL-17 was scarcely expressed and secreted in the untreated control untreat group.The cells expression of IL-17 were significantly different among the four groups (F=160.549, P＜0.01).The cells expressing of IL-17 were higher in the miR-155 mimics group (39.86±4.62)％ than those at the miR-155 inhibitor group (22.02±2.81)％, P＜0.01) and in the treated control treat group [(19.44±1.49)％, P＜0.01].The level of IL-17 was also significantly different among the four groups (F=260.813, P＜0.01).The level of IL-17 was higher in the miR-155 mimics group [(1 509±136) pg/ml] than that in the miR-155 inhibitor group [(923± 42) pg/ml, P＜0.01);and in the treated control group [(767±94) pg/ml, P＜0.01).The expression of miR-155 (12.53±0.80 vs 1.78±0.14, 7.16±0.62, 6.47±0.92, P＜0.01) and IL-17A mRNA (46.55±6.71 vs 1.01±0.19,15.62±1.26, 14.20±2.73, P＜0.01) was significantly higher than that in the other three groups, while the expression of Ets-1 mRNA was significantly lower (0.66±0.10 vs 1.19±0.04, 1.01±0.16, 1.37±0.27, P＜0.01).si-Ets-2 was screened because it markedly inhibited the expression of Ets-1 mRNA among the three designed siRNAs.The expression of IL-17A mRNA was higher (17.19±3.58 vs 10.08±0.76, t=-3.361, P=0.028) and the expression of Ets-1 mRNA was lower (0.27±0.01 vs 0.74±0.03, t=-30.275, P＜0.01) in si-Ets-2 group than that in si-Con group when si-Ets-2 or si-Con was co-transfected with miR-155 mimics or inhibitor lentiviral vectors.The expression of Ets-1 protein was lower in si-Ets-2 group than that in si-Con group by Western blotting and the decrease was markedly obvious in the miR-155 mimics group.Conclusion miR-155 can induce CD4+T cells to differentiate into Th17 cells by inhibiting the gene expression of Ets-1.

Preliminary study on peroxisome proliferator activated receptor gamma of lymphocytes in patients with asthma / 重庆医学

Objective To explore the peroxisome proliferator activated receptor gamma (PPARγ) expression characteristic of lymphocytes in patients with asthma .Methods Collected blood samples from healthy subjects (health control group) and asthmatic patients(asthmatic group) before treatment ,2 and 4 days after treatment .Expression levels of PPARγtested with Q-PCR .Analyzed eosinophil percentage of induced sputum ,IL-5 concentration in sputum supernatant measured with ELISA kits .Results Compared with healthy control group ,the eosinophil percentage and IL-5 concentration were higher in asthmatic group before treatment ;meanwhile the expression level of PPARγwas at the lowest .After treatment ,PPARγgradually increased accompanied with eosino-phil percentage and IL-5 concentration gradually decreased .Conclusion Asthmatic patients had a lowest PPARγ expression level . Their recovery perhaps attributed to the up-regulation of PPARγin lymphocytes .The anti-inflammatory effect of PPARγachieved might be via inhibiting the function of Th2 cells .

Residue and Degradation of Cyantraniliprole and Its Main Metabolite in Pepper and Soil / 分析化学

An analytical method based on ultra performance liquid chromatography-tandem mass spectrometry was developed for the determination of cyantraniliprole and its main metabolite J9 Z38 residues in pepper and soil. The fate of cyantraniliprole and J9Z38 in pepper and soil was also evaluated. The target compounds were extracted with acetonitrile, cleaned up by C18 cartridge, and further analyzed by gradient ultra performance liquid chromatography-tandem mass spectrometry with electrospray ionization in positive mode ( ESI﹢) using a UPLC BEH C18 Column. The method was validated using fortified pepper and soil. Intra-day mean recoveries of cyantraniliprole and J9Z38 at three spiked levels (0. 01, 0. 10 and 1. 00 mg/kg) ranged from 88. 6% to 105 . 7% with relative standard deviations of 3 . 8%-15 . 1%. Inter-day mean recoveries of cyantraniliprole and J9 Z38 were found between 91 . 4% and 105 . 3% with relative standard deviations of 4 . 9%-12 . 3% at three spiked levels. Limits of quantification ( LOQs) of cyantraniliprole and J9Z38 were 0. 1 and 0. 2 μg/kg, respectively. Linear calibration functions with correlation coefficients of r>0. 9992 were obtained in the concentration range of 2. 0-128. 0 μg/L. This method was applied to the analysis of cyantraniliprole and J9Z38 residues in real pepper and soil samples selected from field. The results of the residue dynamic experiment showed that the half-life of cyantraniliprole ranged from 9 . 2 to 11 . 2 days in pepper and from 9 . 2 to 20. 8 days in soil. While, the residues of J9Z38 in pepper were below LOQ, and the half-life of J9Z38 in soil was 9. 4 days. The degradation speed of cyantraniliprole increased with the increase of the precipitation.

AFLP analysis of genetic diversity of Alpinia officinarum / 中国中药杂志

<p><b>OBJECTIVE</b>To explore the genetic diversity and relationship of different Alpinia officinarum germplasm.</p><p><b>METHOD</b>Amplified fragment length polymorphism (AFLP) markers were developed to analyze genetic polymorphism in A. officinarun from eight resources. The amplified fragments were used as primary matrix with NTSYSpc-2.11F software to analyze the similarity between the A. officinarum germplasm and to construct the genetic phylogenetic tree.</p><p><b>RESULT</b>A total of 1,120 fragments were genotyped using AFLP with eight prime combinations. Analysis identified 1,044 polymorphic fragments, accounting for 92.57% of the total detected variation. Genetic phylogenetic tree analysis indicates that three categories can be divided among the eight resources of A. officinanrum.</p><p><b>CONCLUSION</b>Significant polymorphism and genetic diversity can be observed among A. officinarum germplasm resources.</p>

Identification of differentially expressed genes in different anomalies of Glycyrrhiza uralensis by cDNA-AFLP method / 中国中药杂志

<p><b>OBJECTIVE</b>To understand the molecular mechanism in genetic variation within species, and identify the differentially expressed genes among 8 anomalies of Glycyrrhiza uralensis under the same genetic background and cultivated environment.</p><p><b>METHOD</b>The differentially expressed genes in roots of different anomalies of G. uralensis in vigorous growing stage were identified by cDNA-AFLP and confirmed by reverse-Northern hybridization. Their functions were inferred through bioinformatics method.</p><p><b>RESULTS</b>Fourteen differentially expressed genes were identified, among which, the function of 6 were unknown, and the rest were involved in antibacterium, regulating metabolism, enhancing the heat resistance ability, increasing the nitrogen fixation ability, and other bioprocesses.</p><p><b>CONCLUSION</b>For the first time, the differentially expressed genes were cloned from the different anomalies of G. uralensis. This provided the basis for the screening of fine anomalies and varieties, and the research of functional genomics.</p>

Evaluation of photodynamic therapy of skin cancers with partial differential alpha-aminolevulinic acid / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To use partial differential -aminolevulinic acid induced photodynamic therapy (ALA-PDT) increasingly in treating skin cancers and other diseases in many countries and to explore the efficacy of ALA-PDT for skin cancers in China.</p><p><b>METHODS</b>Eighty-eight patients, including 34 cases of basal cell carcinoma (BCC), 32 cases of squamous cell carcinoma (SCC), two cases of basal-squamous cell carcinoma (BSCC), one case of verrucuous carcinoma, nine cases of Bowen disease, two cases Paget disease of the nipple and eight cases of extramammary Paget disease, were treated by the partial differential alpha-aminolevulinic acid induced photodynamic therapy first in China from 1997 to 2000.</p><p><b>RESULTS</b>All BCC, including 11 cases of superficial lesions and 29 solid lesions, achieved complete reaction (CR) by 1-4 times of the ALA-PDT. Except one patient with adenoid SCC (grade III), all SCC (grade I and grade II) patients achieved complete remission by 3-6 times of ALA-PDT. All Bowen diseases achieved complete reaction by 1-4 times. Although for Paget diseases it could not cure the disease simply by ALA-PDT, it could control the symptoms. The recurrence rates were 11% (4/34) for BCC, and 22% (7/32) for SCC by following up 1-3 years after the therapy. The continuous therapy is still effective.</p><p><b>CONCLUSIONS</b>ALA-PDT is an effective, non-traumatic treatment for patients with BCC, SCC, Bowen and Paget diseases. It is especially suitable for older and weaker patients or those who are not tolerable to other therapies. It also has a unique advantage for tumors in specific anatomical areas. It is a new alternative modality for skin cancer therapies.</p>

Production of puerarin and other isoflavones in cell suspension cultures in leaves of Pueraria lobata seedling / 中草药

Object To establish cell suspension culture system in leaves of seedling of Pueraria lobata (Willd.) Ohwi Methods The callus induced from leaf explants of P.lobata seedling were suspended in liquid medium. The time courses of cell growth and yields of puerarin and isoflavones were studied Results The log phase of the cell cultures' growth was from 1st d through 5th d. The dry weight accumulation reached highest on 8th d. The production of puerarin was highly coupled with the cell growth. The yield of puerarin reached highest on 8th d. However, the accumulation of total isoflavones was different from that of puerarin. The yield of total isoflaones reached the highest on 10th d Conclusion The yield of total isoflavones in cell suspension culture system is higher than that in wild plants, while the yield of puerarin is lower than that in wild plants