高分子聚合物HFMC注入猕猴输精管后附睾的远期组织病理学变化

本实验采用７只成年雄性猕猴,经双侧阴囊上方切口暴露输精管,由近附睾端向远侧注射高分子聚合物ＨＦＭＣ,剂量分别为３０ｍｇ／侧（１只）,６０ｍｇ／侧（３只）,１００ｍｇ／侧（３只）,分别于术后２．５及３．５年处死动物,取附睾组织进行光镜及电镜观察。结果表明：注射不同剂量ＨＦＭＣ２．５年后,动物附睾头、体、属各种光镜观察所见主要改变为上皮细胞局灶性轻度水样变性,少许上皮细胞脱落,个别管腔扩张,局部间质少     

黄芪复合体（豆科）核型研究补充材料

本文首次报道了民和黄芪的染色体数目及核型。发现该种与黄芪复合体其它类群的染色体数目相同,但核型有别,其核型公式为２ｎ＝１６＝８ｍ（２ＳＡＴ）＋８ｓｍ。这种核型变异与它的形态变异一致。在黄芪复合体内,每一类群的染色体至少具１对随体,且附着在最后１对染色体的短臂上（除蒙古黄芪具２对随体外）,而Ｔｏｈ（１９７１）报道采自Ｋｙｕｎｇｉ和Ｍｔ．Ｈａｒｌａ膜荚黄芪和高山膜荚黄芪（新拟）的染色体不具随体,可能观察有误。     

Plasma microRNA profiling highlights miR-1260b and miR-720 as novel diagnostic and prognostic biomarkers of esophageal squamous cell carcinoma

This paper was designed to explore the value of miRNAs as diagnostic biomarkers that may facilitate the early detection of esophageal squamous cell carcinoma (ESCC). Plasma miRNA profiles were defined via an array-based approach using samples from ESCC patients and healthy controls (n=5 each). Differentially expressed miRNAs in these samples were validated via qPCR in ESCC patients (n=96) and healthy controls (n=51), and the relationship between ESCC patient plasma miR-1260b and miR-720 levels and clinicopathological characteristics were additionally examined. In total, 12 plasma miRNAs that were differentially expressed between ESCC patients and healthy controls were identified via miRNA. Six of these miRNAs were subsequently validated, revealing that both miR-1260b and miR-720 were significantly differentially abundant in ESCC patients and controls, with miR-1260b being significantly upregulated in ESCC patients relative to controls (2.24, 1.41 respectively, P<0.001), while the opposite was observed with respect to miR-720 (0.66, 2.27 respectively, P=0.001). The use of both miR-720 and miR-1260b as a combined diagnostic tool was highly efficacious, yielding an AUC of 0.814, a sensitivity of 86.3%, and a specificity of 73.2% as a means of detecting ESCC patients. Elevated plasma miR-1260b level was also associated with a poorer patient prognosis when compared to patients with a low plasma miRNA level (P=0.021). This study has successfully developed a plasma miRNA biomarker signature of ESCC that may offer value as a diagnostic or prognostic tool when evaluating patients with ESCC.     

Stability of actin cytoskeleton and PKC-delta binding to actin regulate NKCC1 function in airway epithelial cells

Activation of airwayepithelial Na-K-2Cl cotransporter (NKCC)1 requires increased activityof protein kinase C (PKC)-, which localizes predominantly to theactin cytoskeleton. Prompted by reports of a role for actin in NKCC1function, we studied a signaling mechanism linking NKCC1 and PKC.Stabilization of actin polymerization with jasplakinolide increasedactivity of NKCC1, whereas inhibition of actin polymerization withlatrunculin B prevented hormonal activation of NKCC1. Protein-proteininteractions among NKCC1, actin, and PKC- were verified by Westernblot analysis of immunoprecipitated proteins. PKC- was detected inimmunoprecipitates of NKCC1 and vice versa. Actin was also detected inimmunoprecipitates of NKCC1 and PKC-. Pulldown of endogenous actinrevealed the presence of NKCC1 and PKC-. Binding of recombinantPKC- to NKCC1 was not detected in overlay assays. Rather, activatedPKC- bound to actin, and this interaction was prevented by a peptideencoding C2, a C2-like domain based on the amino acid sequence ofPKC-. C2 also blocked stimulation of NKCC1 function bymethoxamine. Immunofluorescence and confocal microscopy revealedPKC- in the cytosol and cell periphery. Merged images of cellsstained for actin and PKC- indicated colocalization of PKC- andactin at the cell periphery. The results indicate that actin iscritical for the activation of NKCC1 through a direct interaction with PKC-.

     

Transplantation of GFP-expressing Blastomeres for Live Imaging of Retinal and Brain Development in Chimeric Zebrafish Embryos

Cells change extensively in their locations and property during embryogenesis. These changes are regulated by the interactions between the cells and their environment. Chimeric embryos, which are composed of cells of different genetic background, are great tools to study the cell-cell interactions mediated by genes of interest. The embryonic transparency of zebrafish at early developmental stages permits direct visualization of the morphogenesis of tissues and organs at the cellular level. Here, we demonstrate a protocol to generate chimeric retinas and brains in zebrafish embryos and to perform live imaging of the donor cells. The protocol covers the preparation of transplantation needles, the transplantation of GFP-expressing donor blastomeres to GFP-negative hosts, and the examination of donor cell behavior under live confocal microscopy. With slight modifications, this protocol can also be used to study the embryonic development of other tissues and organs in zebrafish. The advantages of using GFP to label donor cells are also discussed. Download video file.^{(95M, mp4)}     

Possible role of Pax-6 in promoting breast cancer cell proliferation and tumorigenesis

Pax 6, a member of the paired box (Pax) family, has been implicated in oncogenesis. However, its therapeutic potential has been never examined in breast cancer. To explore the role of Pax6 in breast cancer development, a lentivirus based short hairpin RNA (shRNA) delivery system was used to knockdown Pax6 expression in estrogen receptor (ER)-positive (MCF-7) and ER-negative (MDA-MB-231) breast cancer cells. Effect of Pax6 silencing on breast cancer cell proliferation and tumorigenesis was analyzed. Pax6-RNAi-lentivirus infection remarkably downregulated the expression levels of Pax6 mRNA and protein in MCF-7 and MDA-MB-231 cells. Accordingly, the cell viability, DNA synthesis, and colony formation were strongly suppressed, and the tumorigenesis in xenograft nude mice was significantly inhibited. Moreover, tumor cells were arrested at G0/G1 phase after Pax6 was knocked down. Pax6 facilitates important regulatory roles in breast cancer cell proliferation and tumor progression, and could serve as a diagnostic marker for clinical investigation.     

Two CYP71AJ enzymes function as psoralen synthase and angelicin synthase in the biosynthesis of furanocoumarins in Peucedanum praeruptorum Dunn

Plant Molecular Biology - Psoralen synthase and angelicin synthase responsible for the formation of psoralen and angelicin in Peucedanum praeruptorum Dunn were identified and functionally...     

Variation of bacterial and fungal community structures in the rhizosphere of hybrid and standard rice cultivars and linkage to CO2 flux

A field experiment was conducted with cultivation of hybrid and conventional cultivars in a rice paddy from China. Rhizosphere soil was sampled and CO(2) flux was measured at tillering (S1), grain filling (S2) and ripening (S3) across the growth stages. Microbial community structure, abundance and activity were analyzed using a combination of functional (enzymes) and denaturing gradient gel electrophoresis (DGGE) and real-time PCR molecular approaches. Invertase and urease activities, total microbial biomass carbon, bacterial 16S rRNA and fungal internal transcribed spacer rRNA gene copies were found to be the highest at S2 under both cultivars, being greater under the hybrid cultivar than under the conventional cultivar across the stages. Moreover, the CO(2) flux was 11%, 16% and 25% higher under the hybrid cultivar than under the conventional cultivar at S1, S2 and S3, respectively. Principal component analyses of the PCR-DGGE profile revealed a significant difference between conventional and hybrid cultivars across growth stages. Sequencing DGGE bands of the bacterial 16S rRNA gene showed that a particular bacterial group of Alphaproteobacteria was enhanced and several distinct operational taxonomic units markedly resembled Ascomycota under the hybrid cultivar. These illustrate a significant selection of a particular group of bacteria and fungi of the hybrid cultivar. However, the potential impacts of these cultivar effects in soil C and N cycling deserve further field studies.     

The mitochondrial genome of Polylabris halichoeres (Monogenea: Microcotylidae)

This study presents the complete mitochondrial (mt) genome of Polylabris halichoeres, which is the largest mt genome sequenced of monogeneans so far and the second complete sequence after Microcotyle sebastis from the Microcotylidae. It is basically similar to that of M. sebastis, with the exception of a high level of gene rearrangement located between trnC and trnL((UUR)), a translocation of trnM and trnH, as well as a highly repetitive region (HRR) in the large non-coding region (NCR). We also find a series of trnI pseudogenes (ΨI) and one unknown short open reading frame (ORF) in the large NCR. Although the ORF cannot be unambiguously regarded as an atp8 gene, we cannot rule out the possibility that it has other functional importance, but it need further study in the future.