Analysis of lycopodiastrum casuarinoides transcriptome discovers lycodine-type alkaloid biosynthetic genes and genetic markers / 中国药理学与毒理学杂志

OBJECTIVE o facilitate the basic acquaintance with the bioactive lycodine-type alka-loids biosynthetic pathways, we conducted the transcriptome analysis of L. casuarinoides by illumina sequencing.METHODS The plant of L.casuarinoides was collected and was subjected to RNA isolation, cDNA library construction, and illumina sequencing before bioinformatics analysis. After sequencing, the clean reads were obtained for de novo assembly by using Trinity software,and then further processed with TGICL sequencing clustering software to generate unigenes,The unigenes are aligned by Blast X alignment to six public protein database. In addition, all unigenes are functionally annotated by GO, KEGG and characterized putative genes involved in lycopodium alkaloids biosynthesis. RESULTS In total, 124, 524 high-quality unigenes were obtained with an average sequence length of 601 bp. Among the L. casuarinoides transcripts, 61,304 showed significant similarity (E-value<1 e-5) to the known proteins in the public database.Among the total 124 524 unigenes,47,538 open reading frame (ORFs) were predicted. Based on the bioinformatics analysis, all possible enzymes involved in the Lycodine-type alkaloids biosynthetic pathway of L. casuarinoides were identified, including primary amine oxidase (PAO), and Malonly-CoA decarboxylase. In addition, a total of 64 putative cytochrome P450 (CYP450) and 827 transcription factors were selected as the candidates of Lycodine-type alka-loids modifiers. Furthermore, a total of 13 352 simple sequence repeats (SSRs) were identified from the 124, 524 unigenes, of which dinucleotide motifs AG/CT (50.1%), were the most abundant. CONCLU-SION This transcriptome analysis of L.casuarinoides,provides many valuable candidate genes involv-ing in the biosynthesis of novel secondary metabolites but also lays the foundation for genetic diversity analysis via SSRs markers in L.casuarinoides.

Mechanism of laparoscopic adjustable gastric banding in the treatment of obesity with type 2 diabetes mellitus / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To explore the mechanism of laparoscopic adjustable gastric banding (LAGB) in the treatment of obese patients with type 2 diabetes mellitus (T2DM).</p><p><b>METHODS</b>A total of 20 patients with obesity and T2DM were treated with LAGB. During the postoperative 1, 3, 6, 9, 12 months, the body weight changes were monitored and body mass indices (BMI) were calculated. The serum levels of leptin, GLP-1, and ghrelin were examined preoperatively and 1, 3, 6, 9, 12 months after LAGB using enzyme-linked-immunosorbent assay (ELISA). At the same time, the fasting serum insulin (FINS), C-peptide, glycated hemoglobin (HbA1c) levels were examined by electrochemiluminescence and the level of fasting blood glucose (FBG) was tested with oxidase test.</p><p><b>RESULTS</b>At postoperatively 12 months, all the 20 patients lost weight. The mean body weight decreased from (108 + or - 18) kg to (71 + or - 16) kg (P<0.05) and BMI decreased from 38 + or - 5 to 29 + or - 6 (P<0.05). The HOMA-IR decreased from (12.8 + or - 7.4) to (3.4 + or - 2.0) (P<0.01). The serum ghrelin level increased from (7.8 + or - 1.9) microg/L to (11.6 + or - 2.6) microg/L (P<0.01). The serum leptin level declined from (24.9 + or - 13.7) microg/L to(12.9 + or - 5.1) microg/L (P<0.01). The serum GLP-1 level increased from (0.58 + or - 0.12) microg/L to(0.80 + or - 0.06) microg/L (P<0.01). After LAGB, there were positive correlations between serum leptin level and FBG, FINS, HbA1c,and C-peptide level. Serum ghrelin and GLP-1 were negatively correlated with FBG, FINS, HbA1c,C-peptide.</p><p><b>CONCLUSIONS</b>LAGB is effective in treatment of obesity patients with T2DM. The mechanism may be associated with the increase of serum GLP-1 and ghrelin and the decrease of serum leptin and insulin resistance.</p>

Immune regulation effect of rat dendritic cells phagocytosing photochemotherapy-treated allogeneic cells on syngeneic T cells / 中国实验血液学杂志

The aim of this study was to investigate the immune regulatory effect of dendritic cells phagocytosing photochemotherapy-treated allogeneic spleen lymphocytes on syngeneic T cells. DA rat spleen lymphocytes were treated with 8-methoxypsoralen plus UVA irradiation (PUVA). LEW rat bone marrow-derived DCs were co-cultured with PUVA-treated DA spleen lymphocytes (PUVA-SP), and the surface markers (MHC-II, CD86 and CD40) of treated DC were detected by flow cytometry. CFSE-labeled PUVA SP were incubated with LEW DCs and the phagocytosis of DCs on PUVA-SP was observed by using fluorescent microscope. The ability of DC phagocytosing allogeneic PUVA-SP (PUVA-SP DC) to stimulate the proliferation of LEW T cells was analyzed by mixed leukocyte reactions (MLR). The production of IL-4, IL-10, IL-2, IFN-gamma in MLR culture supernatant was determined by luminex method. The results indicated that the PUVA treatment effectively induced early apoptosis of DA rat spleen lymphocytes. After co-culture, DC efficiently phagocytosed allogeneic PUVA-SP and still maintained an immature phenotype with low levels of MHC II, CD40 and CD86. PUVA-SP DC induced LEW T cell hyporesponsiveness to DA rat antigen, and led to skewing of T cell cytokine expression toward Th2 (IL-10 and IL-4). It is concluded that the PUVA-SP DC effectively down-regulate T cell response to alloantigen and induce Th2 immune deviation in vitro.