[Research progress on liposome and nanomicelle targeted drug delivery system across blood-brain barrier].

The blood-brain barrier(BBB), a protective barrier between brain tissues and brain capillaries, can prevent drugs from entering the brain tissues to exert the effect, which greatly increases the difficulty in treating brain diseases. The drug delivery system across the BBB can allow efficient drug delivery across the BBB by virtue of carriers and formulations, thereby enhancing the therapeutic effect of drugs on brain tissue diseases. Liposomes and micelles have been extensively studied with advances in the targeted therapy across the BBB for the brain due to their unique structures and drug delivery advantages. This study summarized the research status of liposome and micelle drug delivery systems across the BBB based on the literature in recent years and analyzed their application advantages and mechanism in terms of trans-BBB capability, targeting, and safety. Moreover, the problems and possible countermeasures in the research on trans-BBB liposomes and micelles were discussed according to the current clinical translation, which may provide refe-rences and ideas for the development of trans-BBB targeted nano-drugs.

Prevalence, morphology, and molecular characteristics of Sarcocystis spp. in domestic goats (Capra hircus) from Kunming, China.

Despite the importance of worldwide goat production, little is known about the prevalence of Sarcocystis spp. in domestic goats (Capra hircus) in China. The aims of the present study were to determine prevalence of Sarcocystis spp. in domestic goats in Kunming, China, as well as to identify parasite species based on morphological characteristics and DNA sequence analysis. Only microscopic sarcocysts of Sarcocystis spp. were detected in 174 of 225 goats (77.3 %). By light and transmission electron microscopy, two species, i.e., Sarcocystis capracanis and Sarcocystis hircicanis, were identified. Two sarcocysts from each of the two species were randomly selected for DNA extraction; the 18S rRNA gene (18S rRNA), the 28S rRNA gene (28S rRNA), and the mitochondrial cytochrome c oxidase subunit 1 (cox1) were amplified by the polymerase chain reaction (PCR) and subsequently sequenced. The results were compared with other previously sequenced Sarcocystis species retrieved from GenBank. There was little sequence variation between two isolates of the same species. S. capracanis was most closely related with Sarcocystis tenella; 18S rRNA, 28S rRNA, and mitochondrial cox1 sequences shared identities of 95.7-99.1, 95.3, and 92.3-93.2 % with those of S. tenella, respectively. Thus, mitochondrial cox1 sequences seem to perform better than 18S rRNA sequences or 28S rRNA sequences for identification of the two species. S. hircicanis was most closely related to Sarcocystis arieticanis, i.e., 18S rRNA and 28S rRNA sequences of the former species shared 97.2-97.4 and 95.6-96.1 % identities with those of latter, respectively. Phylogenetic analysis inferred from the three genetic markers yielded similar results and indicated the two species were within a group of Sarcocystis species with canines as known, or presumed, definitive hosts.

Sarcocystis clethrionomyelaphis Matuschka, 1986 (Apicomplexa: Sarcocystidae) infecting the large oriental vole Eothenomys miletus (Thomas) (Cricetidae: Microtinae) and its phylogenetic relationships with other species of Sarcocystis Lankester, 1882.

Sarcocystis clethrionomyelaphis Matuschka, 1986 was first identified in skeletal muscles of 47 (75.8%) of 62 large oriental voles Eothenomys miletus (Thomas) captured between March 2012 and May 2014 in Anning Prefecture of Yunnan Province (China). Sarcocyst walls were thick and possessed villous protrusions measuring 3.5-5.5 µm in length. Beauty rat snakes Elaphe taeniura (Cope) fed sarcocysts of the species shed sporulated oöcysts measuring 13-18×9-13 (16×12) µm with a prepatent period of 16 to 17 days. Phylogenetic analysis based on 18S rRNA gene sequences revealed a close relationship between S. clethrionomyelaphis and other colubrid-transmitted species of Sarcocystis Lankester, 1882. This is the first report identifying S. clethrionomyelaphis from its natural intermediate host.

Increased CAP37 Expression in Stable Chronic Obstructive Pulmonary Disease.

OBJECTIVE: Cationic antimicrobial protein of 37 kDa (CAP37), a neutrophil-derived protein originally identified for its antimicrobial activity, is now known to have many regulatory effects on host cells. However, its role in the pathogenesis of chronic obstructive pulmonary disease (COPD) has not been studied. We therefore investigated the expression of CAP37 in COPD and its effects on airway structural cells, including bronchial epithelial cells, smooth muscle cells, and fibroblasts. METHODS: CAP37 was detected in the lung tissue, sputum, and plasma of COPD patients and the control subjects, as well as in the neutrophils stimulated by cigarette smoke extract (CSE). BEAS-2B cells, human bronchial smooth muscle cells (HBSMCs), and MRC-5 cells were treated with CAP37 or an anti-CAP37 antibody plus CAP37. Interleukin (IL)-6 and IL-8 were detected in the BEAS-2B cells. The cell proliferation was analyzed in the HBSMCs. Collagens were also detected in the MRC-5 cells. RESULTS: The expression of CAP37 was increased in the lung tissue and sputum supernatant of the COPD patients compared with the control subjects. The sputum supernatant CAP37 levels were inversely correlated with the forced expiratory volume in the first second percentage predicted in COPD. CAP37 was induced by CSE stimulation in the peripheral blood neutrophils from healthy non-smokers. CAP37 induced expression of IL-6 and IL-8 in BEAS-2B cells, and collagen expression of lung fibroblasts (MRC-5 cells). However, CAP37 did not significantly alter the proliferation of the HBSMCs. CONCLUSION: Our findings indicated that neutrophil-derived CAP37 may be involved in airway inflammation and fibrosis in COPD via affecting the bronchial epithelial cells, and fibroblasts, thus suggesting a possible role of CAP37 in the development and progression of COPD.

[Construction of vectors for expression of cleavable tandem repeat Thanatin fusion protein in plants].

Thanatin, a 21-residue antimicrobial peptide, is an inducible insect peptide with a broad range of activity against bacteria and fungi. It can improve the expression level of the antimicrobial peptide in plants to express the peptide as tandem repeat fusion protein containing multiple Thanatin copies. However, the fusion protein has no antimicrobial activity. To make the fusion protein automatically break into single Thanatin unit with antimicrobial activity, the fused Thanatin was spaced by a linker peptide, which was cleavable in vivo. The soybean (Glysine max L.) chitinase signal peptide was fused to the N end of the fusion protein to induce the antimicrobial peptide accumulated in intercellular space. To construct the vectors for expression of the fusion protein, the overlapped primers were used to clone the antimicrobial peptide gene and the co-adhesive end restriction and ligation strategy was used to add the repeat unit one by one. Vectors containing 1 to 5 repeat units of Thanatin were constructed respectively. These vectors were being used to transform plants to improve plant disease resistance.

[Raising fat content in transgenic rice by anti-PEP gene transformation].

Gene for phosphoenolpyruvate carboxylase (PEPCase) was incorporated in antisense orientation into genome of japonica rice variety Xiushui 11 through Agrobacterium tumefaciens-mediated transformation of mature seeds-derived embryogenic calli, and a total of 95 transgenic plants were regenerated, which were confirmed by histochemical detection of GUS activity (Fig.2), PCR assay (Fig.3) and Southern hybridization analysis (Fig.4). The transgene was inherited by the T1 progeny and Mendelian 3:1 segregation ratio was observed in most transgenic lines. It was found that the fat content of hulled grains of T2 transgenic lines was 0.37 +/- 0.12 per cent higher than that of the control, most of which were statistically very significant (Table 2).

Highly efficient transformation and plant regeneration of tall fescue mediated by Agrobacterium tumefaciens.

An efficient and reproducible system has been developed for the production of transgenic plants in tall fescue (Festuca arundinacea Schreb.) using A. tumefaciens-mediated transformation. Two-months-old suspension cultures served as excellent explants for transformation. The explants were inoculated with an A. tumefaciens strain EHA105 carrying a plasmid pDBA121 containing genes for hygromycin phosphotransferase (hpt) and phosphinotricin acetyltransferase (bar). The commercial herbicide Basta was used as a selective agent. Inclusion of acetosyringone (ACS) 20 mg/L in the co-culture medium led to an increase in transformation efficiency. The efficiency of transformation was highly dependent on the genotype, the explant, the culture medium and selective agent used. Tall fescue transformation efficiency is 2.85-10.9 plants per gram fresh weight (FW) of suspension cultures. This is much higher than the corresponding values reported before (2-5 plants). So far more than 300 transgenic plants have been obtained, the fertility of some transgenic plants had decreased. Stable integration and high expression of the transgenes were confirmed by PCR analysis and Southern blot hybridization or herbicide Basta spraying test.