Exploring the Terminal Pathway of Sex Pheromone Biosynthesis and Metabolism in the Silkworm.

Sex pheromones are vital to sexual communication and reproduction in insects. Although some key enzymes in pheromone production have been well studied, information on genes involved in the terminal pathway is limited. The domestic silkworm employs a pheromone blend containing (E,Z)-10,12-hexadecadienol (bombykol) and analogous (E,Z)-10,12-hexadecadienal (bombykal); whereas, its wild ancestor B. mandarina uses only bombykol. The two closely related moths might be a good model for exploring the genes involved in aldehyde pheromone synthesis and metabolism. By deep sequencing and analyzing the sex pheromone gland (PG) transcriptomes; we identified 116 candidate genes that may be related to pheromone biosynthesis, metabolism, and chemoreception. Spatiotemporal expression profiles and differentially expressed analysis revealed that four alcohol oxidases (BmorAO1; 2; 3; and 4); one aldehyde reductase (BmorAR1); and one aldehyde oxidase (BmorAOX5) might be involved in the terminal pathway. Phylogenetic analysis showed that, except for BmorAO3 and MsexAO3, AOs did not show a conversed orthologous relationship among moths; whereas, ARs and AOXs were phylogenetically conserved. This study provides crucial candidates for further functional elucidation, and which may be utilized as potential targets to disrupt sexual communication in other moth pests.

Emerging Insights Into the Role of Epigenetics and Gut Microbiome in the Pathogenesis of Graves' Ophthalmopathy.

Graves' Ophthalmopathy (GO) is an organ-specific autoimmune disease that is often characterized by infiltration of orbital tissues and is considered as the most common extra-thyroid manifestation of Graves' disease (GD). Although genetic susceptibility has been found to be critical for the phenotype of GO, the associated risk alleles in a single gene are generally insufficient to cause the disease. Accruing evidence has shown that epigenetic disorders can act as the potentially missing link between genetic risk and clinically significant disease development. Abnormal epigenetic modifications can lead to pro-inflammatory cascades and activation of orbital fibroblasts (OFs) by promoting the various inflammatory response pathways and regulating the diverse signaling molecules that are involved in the fibrogenesis and adipogenesis, thereby leading to the significant expansion of orbital tissues, fibrosis and inflammation infiltration. Additionally, emerging evidence has shown that the gut microbiome can possibly drive the pathogenesis of GO by influencing the secretion of Thyrotropin receptor antibody (TRAb) and T-helper 17 (Th17)/regulatory T cells (Treg) imbalance. This paper describes the latest epigenetic research evidence and progress made in comprehending the mechanisms of GO development, such as DNA methylation, histone modification, non-coding RNAs, and the gut microbiome.

The Antitumor Immunity and Tumor Responses of Chemotherapy with or without DC-CIK for Non-Small-Cell Lung Cancer in China: A Meta-Analysis of 28 Randomized Controlled Trials.

OBJECTIVE: DC-CIK therapy included DC-CIK cells and Ag-DC-CIK cells. To further confirm whether DC-CIK reconstructs the antitumor immunity and improves the tumor responses and reveals its optimal usage and combination with chemotherapy, we systematically reevaluated all the related studies. MATERIALS AND METHODS: All studies about DC-CIK plus chemotherapy for NSCLC were collected from the published and ongoing database as CBM, CNKI, VIP, Wanfang, ISI, Embase, MEDLINE, CENTRAL, WHO-ICTRP, Chi-CTR, and US clinical trials (established on June 2017). We evaluated their methodological bias risk according to the Cochrane evaluation handbook of RCTs (5.1.0), extracted data following the predesigned data extraction form, and synthesized the data using meta-analysis. RESULTS: We included 28 RCTs (phase IV) with 2242 patients, but most trials had unclear bias risk. The SMD and 95% CI of meta-analysis for CD3+ T cells, CD3+ CD4+ T cells, CD3+ CD8+ T cells, CD4+/CD8+ T cell ratio, CIK cells, NK cells, and Treg cells were as follows: 1.85 (1.39 to 2.31), 0.87 (0.65 to 1.10), 1.04 (0.58 to 1.50), 0.75 (0.27 to 1.22), 3.87 (2.48 to 5.25), 1.51 (0.99 to 2.03), and -2.31(-3.84 to -0.79). The RR and 95% CI of meta-analysis for ORR and DCR were as follows: 1.38 (1.24 to 1.54) and 1.27 (1.20 to 1.34). All differences were statistically significant between DC-CIK plus chemotherapy and chemotherapy alone. Subgroup analysis showed that only DC-CIK cells could increase the CD3+T cells, CD3+ CD4+T cells, CD3+ CD8+T cells, and CD4+/CD8+ T cell ratio. In treatment with one cycle or two cycles and combination with NP or GP, DC-CIK could increase the CD4+/CD8+ T cell ratio. All results had good stability. CONCLUSIONS: DC-CIK therapy can simultaneously improve the antitumor immunity and tumor responses. DC-CIK therapy, especially DC-CIK cells, can improve antitumor immunity through increasing the T lymphocyte subsets, CIK cell, and NK cells in peripheral blood. The one cycle to two cycles may be optimal cycle, and the NP or GP may be optimal combination.

Cloning and sequence analysis of L-lysine decarboxylase gene in Huperzia serrata var. longipetiolata / 中草药

Objective: To obtain and analyze the full-length L-lysine decarboxylase (LDC) gene sequence of Huperzia serrata var. longipetiolata and predictively analyze its protein structure on the basis of cloning the coding region of LDC gene from four species of Huperziaceae. The species are Huperzia serrata var. longipetiolata, Phlegariurus minchegensis, Phlegariurus austrosinicus, and Phlegariurus petiolatus. Methods: The LDC coding region sequences were cloned by RT-PCR strategy with the template of total RNA extracted from the leaves. Then the sequences were analyzed by means of BLAST and MEGA 5.0. The full-length of LDC gene sequence of H. serrata var. longipetiolata was obtained by RACE technology. And then the secondary structure and three-dimensional structure of LDC protein were predictively analyzed. Results: The coding region sequences were highly similar to the lysine decarboxylase in the database. And the encoding protein of H. serrata var. longipetiolata was highly similar to the amino acid sequences of H. serrata in NCBI, and with high homology to Selaginella moellendorffii. The full-length LDC gene sequence of H. serrata var. longipetiolata contained 1 266 bp open reading frame and encodes a predicted protein of 403 amino acids. The GenBank accession number for this gene is KF040056. Conclusion: The LDC genes of the four species of Huperziaceae are cloned in this study. The full-length LDC gene sequence of H. serrata var. longipetiolata is obtained and analyzed, and its protein structure is predictively analyzed. The result will provide a foundation for exploring the mechanism of huperzine A biosynthesis in the plants of Huperziaceae.

[Effect of danshen injection on adult cardiac stem cell transplantation in myocardial infarction mice].

OBJECTIVE: To study the effect of combined use of Danshen Injection (Dl) on acute myocardial infarction (AMI) mice undergoing adult cardiac stem cell transplantation. METHODS: Thirty mice were randomly divided into 3 groups, 10 in each group, i.e., the high dose Dl group (at the daily dose of 5 g/kg) , the low dose Dl group (at the daily dose of 0. 5 g/kg), and the model control group. The AMI model was constructed by surgical ligation of the left anterior descending artery, and 2 x 10(6) -3 x 10(6) cardiac stem cells in vitro cultured were injected to the peripheral infracted zone immediately after successful modeling. All mice were administered with Dl by gastrogavage one week before myocardial infarction (Ml) and one week after Ml. Mice were sacrificed one week after Ml. Immunostaining was performed. The microvessel density (MVD) was detected using CD31 . The cell proliferation was detected using Ki67. The myocardial fibrosis degree was detected using Masson staining. The cell apoptosis of peripheral infracted zone was detected by TUNEL. Protein expressions of Akt and phospho-Akt were detected using Western blot. RESULTS: MVD and Ki67 positive cell number increased more in the high dose DI group than in the low dose DI group and the model control group (P < 0.05). The myocardial fibrosis degree and the cell apoptosis of peripheral infracted zone significantly decreased in the high dose DI group than in the low dose DI group and the model control group (P <0. 05). The phospho-Akt expression level significantly increased in the high dose DI group than in the low dose DI group and the model control group (P < 0.05). Compared with the model control group, the total Akt level significantly increased in the high dose and low dose DI groups (P < 0.05). But there was no statistical difference the high dose DI group and the low dose DI group (P > 0.05). CONCLUSION: Treating AMI by adult cardiac stem cell transplantation combined with DI could increase the MVD and cell proliferation, reduce cell apoptosis and fibrosis levels, thus improving the transplantation efficiency of adult cardiac stem cells.

Evidence of selection at melanin synthesis pathway loci during silkworm domestication.

The domesticated silkworm (Bombyx mori) was domesticated from wild silkworm (Bombyx mandarina) more than 5,000 years ago. During domestication, body color between B. mandarina and B. mori changed dramatically. However, the molecular mechanism of the silkworm body color transition is not known. In the present study, we examined within- and between-species nucleotide diversity for eight silkworm melanin synthesis pathway genes, which play a key role in cuticular pigmentation of insects. Our results showed that the genetic diversity of B. mori was significantly lower than that of B. mandarina and 40.7% of the genetic diversity of wild silkworm was lost in domesticated silkworm. We also examined whether position effect exists among melanin synthesis pathway genes in B. mandarina and B. mori. We found that the upstream genes have significantly lower levels of genetic diversity than the downstream genes, supporting a functional constraint hypothesis (FCH) of metabolic pathway, that is, upstream enzymes are under greater selective constraint than downstream enzymes because upstream enzymes participate in biosynthesis of a number of metabolites. We also investigated whether some of the melanin synthesis pathway genes experienced selection during domestication. Neutrality test, coalescent simulation, as well as network and phylogenetic analyses showed that tyrosine hydroxylase (TH) gene was a domestication locus. Sequence analysis further suggested that a putative expression enhancer (Abd-B-binding site) in the intron of TH gene might be disrupted during domestication. TH is the rate-limiting enzyme of melanin synthesis pathway in insects. Real-time polymerase chain reaction assay did show that the relative expression levels of TH gene in B. mori were significantly lower than that in B. mandarina at three different developmental stages, which is consistent with light body color of domesticated silkworm relative to wild silkworm. Therefore, we speculated that expression change of TH gene may contribute to the body color transition from B. mandarina to B. mori. Our results emphasize the exceptional role of gene expression regulation in morphological transition of domesticated animals.

Proteomic analysis of chronic restraint stress-induced Gan (肝)-stagnancy syndrome in rats / 中国结合医学杂志

<p><b>OBJECTIVE</b>To analyze the proteomic characteristics of Gan (肝)-stagnancy syndrome (GSS) by seeking the differential protein in blood and tissues of GSS model rats.</p><p><b>METHODS</b>GSS model rats were established by chronic restraint stress, keeping rats in restrain chamber for 6 h every day for 21 successive days. Their blood and liver samples were collected at the end of experiment for differential protein detection with methods of isoelectrofocusing and polyacrylamide SDS-PAGE, silver staining, and scanning. The gel images were analyzed with Imagemaster 2D Elite software, and the excavated differential protein spots were identified with matrix assistant laser resolving TOF mass spectrometry, Western blot, ELISA, and RT-PCR, respectively.</p><p><b>RESULTS</b>A method for isolating the protein in blood serum and tissues by two-dimensional gel electrophoresis was established and optimized. Six serum proteins and three liver proteins that differentially expressed were identified. The down-regulated differential proteins in serum of GSS model rats were serum albumin precursor, beta 1 globin, antibody against muscle acetylcholine receptor, Ig lambda-2 C region, and transthyretin (TTR), and those in liver tissue were aryl sulfotransferase, enoyl-CoA hydratase, and TTR. TTR down-regulation was found in both serum and liver. Preliminary biological information analysis showed that these differential proteins involved in immune, neuroendocrine, nutrition, and substance metabolism.</p><p><b>CONCLUSION</b>Proteomic analysis of differential proteins showed that TTR, aryl sulfotransferase, and enoyl-CoA hydratase expressions are downregulated in the GSS model rats, suggesting that the susceptibility of cancer could be enhanced by chronic stress.</p>

Effects of bagasse polysaccharide on the immune functions of immunosuppressed mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the effects of bagasse polysaccharide on the immune functions of immunosuppressed mice.</p><p><b>METHODS</b>Immunosuppressed mouse models were established by intraperitoneal injections with cyclophosphamide followed by daily intragastric administration of bagasse polysaccharide. After the treatments, the mice were examined for immune organ weight index, phagocytotic function of the macrophages, delayed type hypersensitivity, serum IgM level following exposure to chicken red blood cells, formation of hemolytic plaques, T cell percentage and lymphocyte transformation.</p><p><b>RESULTS</b>Treatment of the immunosuppressed mice with bagasse polysaccharide at the daily dose of 200 and 400 mg/kg significantly increased the weight of the immune organs, phagocytotic function of the macrophages, delayed type hypersensitivity, serum IgM level against chicken red blood cells, formation of hemolytic plaques, T cell percentage and lymphocyte transformation.</p><p><b>CONCLUSION</b>Bagasse polysaccharide can enhance the immune functions of immunosuppressed mice.</p>

Protective effects of Hongbeiyegen against immunological liver injury in mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the protective effects of Hongbeiyegen (HBYG) against immunological liver injury induced by bacille Calmette-Guerin (BCG) and lipopolysaccharide (LPS).</p><p><b>METHODS</b>Immunological liver injury was induced in rats by BCG and LPS injected via the tail vein. The liver index, thymus index and spleen index were calculated and the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and nitric oxide (NO) and liver homogenate contents of tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) were determined.</p><p><b>RESULTS</b>HBYG significantly improved the liver index, thymus index and spleen index, and reduced the serum levels of ALT, AST and NO, and as the liver homogenate contents of TNF-alpha and IL-1beta.</p><p><b>CONCLUSION</b>HBYG offers obvious protective effects against immunological injury liver in mice.</p>

Therapeutic effect of Hongbeiyegen on alcohol-induced rat hepatic fibrosis / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the therapeutic effect of Hongbeiyegen [the root of Alchornea trewioides(Benth.) Muell.-Arg.] on alcohol-induced liver fibrosis (AF) in rats and explore its mechanism.</p><p><b>METHODS</b>In rats with AF, the serum levels of transforming growth factor beta1 (TGFbeta1) and tissue inhibitor of metalloproteinase-1 (TIMP-1) were detected along with examination of the changes in serum hyaluronic acid (HA), laminin (LN), procolagen type III (PC III), collagen type IV (C IV), glutamic-pyruvic transaminase (ALT) and glutamic-oxalacetic transaminase (AST) levels.</p><p><b>RESULTS</b>Compared with the control group, Hongbeiyegen could significantly reduce the levels of TGFbeta1, TIMP-1, HA, LN, PC III, CIV, ALT and AST in rats with AF.</p><p><b>CONCLUSION</b>Hongbeiyegen can relieve and ameliorate liver fibrosis possibly by inhibiting the expression of TGFbeta1 and TIMP-1.</p>