Identification of long non-coding RNAs and microRNAs involved in anther development in the tropical Camellia oleifera.

BACKGROUND: Explored the molecular science of anther development is important for improving productivity and overall yield of crops. Although the role of regulatory RNAs, including long non-coding RNAs (lncRNAs) and microRNAs (miRNAs), in regulating anther development has been established, their identities and functions in Camellia oleifera, an important industrial crop, have yet not been clearly explored. Here, we report the identification and characterization of genes, lncRNAs and miRNAs during three stages of the tropical C. oleifera anther development by single-molecule real-time sequencing, RNA sequencing and small RNA sequencing, respectively. RESULTS: These stages, viz. the pollen mother cells stage, tetrad stage and uninucleate pollen stage, were identified by analyzing paraffin sections of floral buds during rapid expansion periods. A total of 18,393 transcripts, 414 putative lncRNAs and 372 miRNAs were identified, of which 5,324 genes, 115 lncRNAs, and 44 miRNAs were differentially accumulated across three developmental stages. Of these, 44 and 92 genes were predicted be regulated by 37 and 30 differentially accumulated lncRNAs and miRNAs, respectively. Additionally, 42 differentially accumulated lncRNAs were predicted as targets of 27 miRNAs. Gene ontology enrichment indicated that potential target genes of lncRNAs were enriched in photosystem II, regulation of autophagy and carbohydrate phosphatase activity, which are essential for anther development. Functional annotation of genes targeted by miRNAs indicated that they are relevant to transcription and metabolic processes that play important roles in microspore development. An interaction network was built with 2 lncRNAs, 6 miRNAs and 10 mRNAs. Among these, miR396 and miR156 family were up-regulated, while their targets, genes (GROWTH REGULATING FACTORS and SQUAMOSA PROMOTER BINDING PROTEIN-LIKE genes) and lncRNAs, were down-regulated. Further, the trans-regulated targets of these lncRNAs, like wall-associated kinase2 and phosphomannose isomerase1, are involved in pollen wall formation during anther development. CONCLUSIONS: This study unravels lncRNAs, miRNAs and miRNA-lncRNA-mRNA networks involved in development of anthers of the tropical C. oleifera lays a theoretical foundation for further elucidation of regulatory roles of lncRNAs and miRNAs in anther development.

Physicochemical characterization of the polysaccharide from Bletilla striata: effect of drying method.

The polysaccharide from Bletilla striata, a traditional Chinese herbal medicine, was obtained by different drying techniques: vacuum-drying (BVPS) or vacuum freeze-drying (BFPS). The effect of drying method on the physicochemical properties of the B striata polysaccharide was evaluated using high size exclusion chromatography coupled to multiangle laser light scattering (HPSEC-MALLS), FT-IR and UV spectroscopy, thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), X-ray diffraction (XRD), scanning electron microscopy (SEM) and atomic force microscopy (AFM). The monosaccharide analysis and pH test revealed that the polysaccharide derived from B. striata was a neutral polysaccharide that is composed of glucose and mannose. The solubility and moisture content test's results demonstrated that BFPS was greater than BVPS. The number average molecular weight (Mn) and the computed average molecular weight (Mw) of 99.3% BFPS were 7.297×10(4)g/mol and 9.545×10(4)g/mol, respectively, whereas the Mn and Mw of 97.6% BVPS were 1.218×10(5)g/mol and 1.472×10(5)g/mol, respectively. The FT-IR and UV results indicated that drying technique has little effect on the structure of the polysaccharide. The thermal analysis showed that weight loss event was at 307.85°C and 305.50°C to BVPS and BFPS, respectively. Furthermore, the XRD confirmed that the polysaccharide was the amorphous nature. However, both SEM and AFM images exhibited that the drying technique had a significant impact on the morphology and conformation of the polysaccharide.

Autoimmune pancreatitis: whole-body 18F-FDG PET/CT findings.

PURPOSE: The study aimed to investigate the function of (18)F-fluorodeoxyglucose positron emission tomography/computed tomography ((18)F-FDG PET/CT) in diagnosing of autoimmune pancreatitis (AIP) and whole-body evaluation. METHODS: Seven AIP patients who underwent (18)F-FDG PET/CT systemic examination in our hospital from August 2010 to March 2012 were analyzed retrospectively. Systemic PET/CT routine scanning and pancreatic delayed scanning were performed in all patients. RESULTS: The mean age of 7 AIP patients (6 male and 1 female) was 54.2 years (range from 42 to 71 years). The pancreas showed diffuse enlargement in 6 cases, and segmental enlargement in 1 case. Fluorodeoxyglucose (FDG) uptake was increased in diseased region. The maximum standardized uptake value (SUVmax) was 4.38 ± 0.90 and increased to 5.31 ± 1.08 after delayed scanning, of which small amounts of inflammatory exudate around pancreas was observed in 4 cases. Extrapancreatic lesions (EPLs) were observed in all 7 cases: lymphadenectasis (n = 5), lymphadenectasis with increased FDG uptake (n = 4); associated sialosis with metabolism enlargement (n = 4); associated cholangitis (n = 4); associated interstitial pneumonia (n = 3); inverted "V" shaped high FDG uptake foci in prostate (n = 5). CONCLUSIONS: AIP is a systemic disease. (18)F-FDG PET/CT can exhibit the characteristics of AIP pancreatic lesions, and also better reflect the changes and metabolic characteristics of extrapancreatic organs. It plays a distinct role in diagnosis, differentiating of AIP and whole-body evaluation.

Design and evaluation of a dry coated drug delivery system with floating-pulsatile release.

The objective of this work was to develop and evaluate a floating-pulsatile drug delivery system intended for chronopharmacotherapy. Floating-pulsatile concept was applied to increase the gastric residence of the dosage form having lag phase followed by a burst release. To overcome limitations of various approaches for imparting buoyancy, we generated the system which consisted of three different parts, a core tablet, containing the active ingredient, an erodible outer shell and a top cover buoyant layer. The dry coated tablet consists in a drug-containing core, coated by a hydrophilic erodible polymer which is responsible for a lag phase in the onset of pulsatile release. The buoyant layer, prepared with Methocel K4M, Carbopol 934P and sodium bicarbonate, provides buoyancy to increase the retention of the oral dosage form in the stomach. The effect of the hydrophilic erodible polymer characteristics on the lag time and drug release was investigated. Developed formulations were evaluated for their buoyancy, dissolution and pharmacokinetic, as well gamma-scintigraphically. The results showed that a certain lag time before the drug released generally due to the erosion of the dry coated layer. Floating time was controlled by the quantity and composition of the buoyant layer. Both pharmacokinetic and gamma-scintigraphic data point out the capability of the system of prolonged residence of the tablets in the stomach and releasing drugs after a programmed lag time.

Design and gamma-scintigraphic evaluation of a floating and pulsatile drug delivery system based on an impermeable cylinder.

A blend of floating and pulsatile principles of drug delivery system seems to present the advantage that a drug can be released in the upper GI tract after a definite time period of no drug release. The objective of this study was to develop and evaluate a floating and pulsatile drug delivery system based on an impermeable cylinder. Pulsatile capsule was prepared by sealing the drug tablet and the buoyant material filler inside the impermeable capsule body with erodible plug. The drug delivery system showed typical floating and pulsatile release profile with a lag time followed by a rapid release phase. The lag time prior to the pulsatile drug release correlated well with the erosion properties of plugs and the composition of the plug could be controlled by the weight of the plug. The buoyancy of the whole system depended on the bulk density of the dosage form. Gamma-scintigraphic evaluation in humans was used to establish methodology capable of showing the subsequent in vivo performance of the floating and pulsatile release capsule. Developed formulations showed instantaneous floating with no drug release during the lag time followed by a pulse drug release. From the gamma-scintigraphic results, the pulsatile release capsule we prepared could achieve a rapid release after lag time in vivo, which was longer than that in vitro. The scintigraphic evaluation could confirm qualitatively that the system with in vitro lag time of 4.0 h provided, with relatively high reproducibility, a pulsatile release occurred around 5.0 h after administration.

[Application of positron emission tomography in diagnosis of liver metastases from colorectal cancer].

OBJECTIVE: To evaluate the value of positron emission tomography (PET) in diagnosing liver metastases from colorectal cancer. METHODS: Eighteen patients suspected with liver metastases after resection of colorectal cancer and three patients suspected with other diseases were diagnosed by PET and CT before operation. The result of both diagnostic approaches was compared with the result of exploratory operation. RESULTS: Seventeen of 18 patients were confirmed as liver metastases after resection of colorectal cancer, in whom 14 patients had other synchronous metastases outside liver metastasis including lung metastasis (n= 2), abdominal wall metastasis (n= 2 ), bone metastasis (n= 1), peritoneal cavity lymph nodes metastasis (n= 6), mediastinal lymph nodes (n= 2), virchow lymph node metastasis (n= 1). One patient with negative PET diagnosis was still alive with cancer- free after 1 year followed- up. Three patients suspected with other diseases were also diagnosed as liver metastases from colorectal cancer by PET. CONCLUSION: PET has higher sensitivity in diagnosing liver metastases or other synchronous metastases after resection of colorectal cancer, which suggests that PET can guide the determination of second operative surgery for liver metastases after resection of colorectal cancer.