Common microRNA⁻mRNA Interactions in Different Newcastle Disease Virus-Infected Chicken Embryonic Visceral Tissues.

To investigate the roles and explore the altered expression of microRNAs (miRNAs) and mRNAs in chicken embryos in response to Newcastle disease virus (NDV) infection, deep sequencing was performed. Then, a conjoint analysis of small RNA-seq and mRNA-seq was performed to screen interactional miRNA⁻mRNA pairs during NDV infection. In total, 15 and 17 up- and downregulated miRNAs were identified that potentially targeted 4279 and 6080 mRNAs in NDV-infected chicken embryonic tissues, respectively; in addition, 595 upregulated and 480 downregulated mRNAs were identified. The conjoint analysis of the obtained data identified 1069 miRNA⁻mRNA pairs. Among these pairs, 130 pairs were related to immune or inflammatory responses. The relationship between gga-miR-203a and its target transglutaminase 2 (TGM2) was confirmed using a dual-luciferase reporter system and a real time quantitative polymerase chain reaction (RT-qPCR) assay. Overall, the discovery of miRNAs, mRNAs, and their potential pairing relationships, which may be involved in the regulation of NDV infection, will facilitate our understanding of the complex regulatory relationship between the host and the virus.

Optimization of fatty alcohol biosynthesis pathway for selectively enhanced production of C12/14 and C16/18 fatty alcohols in engineered Escherichia coli.

BACKGROUND: With the increasing stress from oil price and environmental pollution, aroused attention has been paid to the microbial production of chemicals from renewable sources. The C12/14 and C16/18 alcohols are important feedstocks for the production of surfactants and detergents, which are widely used in the most respected consumer detergents, cleaning products and personal care products worldwide. Though bioproduction of fatty alcohols has been carried out in engineered E. coli, several key problems have not been solved in earlier studies, such as the quite low production of C16/18 alcohol, the lack of optimization of the fatty alcohol biosynthesis pathway, and the uncharacterized performance of the engineered strains in scaled-up system. RESULTS: We improved the fatty alcohol production by systematically optimizing the fatty alcohol biosynthesis pathway, mainly targeting three key steps from fatty acyl-acyl carrier proteins (ACPs) to fatty alcohols, which are sequentially catalyzed by thioesterase, acyl-coenzyme A (CoA) synthase and fatty acyl-CoA reductase. By coexpression of thioesterase gene BTE, acyl-CoA synthase gene fadD and fatty acyl-CoA reductase gene acr1, 210.1 mg/L C12/14 alcohol was obtained. A further optimization of expression level of BTE, fadD and acr1 increased the C12/14 alcohol production to 449.2 mg/L, accounting for 75.0% of the total fatty alcohol production (598.6 mg/L). In addition, by coexpression of thioesterase gene 'tesA, acyl-CoA synthase gene fadD and fatty acyl-CoA reductase gene FAR, 101.5 mg/L C16/18 alcohol was obtained, with C16/18 alcohol accounting for 89.2% of the total fatty alcohol production. CONCLUSIONS: To our knowledge, this is the first report on selective production of C12/14 and C16/18 alcohols by microbial fermentation. This work achieved high-specificity production of both C12/14 and C16/18 alcohols. The encouraging 598.6 mg/L of fatty alcohols represents the highest titer reported so far. In addition, the 101.5 mg/L 89.2% C16/18 alcohol suggests an important breakthrough in C16/18 alcohol production. A more detailed optimization of the expression level of fatty alcohol biosynthesis pathway may contribute to a further improvement of fatty alcohol production.

[Impact analysis of atmospheric state for target detection in hyperspectral radiance image].

Target detection based on hyperspectral radiance images can improve data processing efficiency to meet the requirements of real-time processing. However, the spectral radiance acquired by the remote sensor will be affected by the atmosphere. In the present paper, hyperspectral imaging process is simulated to analyze the effects of the changes in atmospheric state on target detection in hyperspectral radiance image. The results show that hyperspectral radiance image can be directly used for target detection, different atmospheric states have little impacts on the RXD detection, whereas the MF detection is dependent on the accuracy of the input spectrum, and good results can only be obtained by the MF detector when the atmospheric states are similar between the radiance spectrum of the target to be detected and the simulated hyperspectral image.

Characterization of chicken IFI35 and its antiviral activity against Newcastle disease virus.

Interferon-induced protein-35 kDa (IFI35) was an antiviral protein induced by interferon (IFN)-γ, which plays an important role in the IFN-mediated antiviral signaling pathway. Here, we cloned and identified IFI35 in the chicken for the first time. Chicken IFI35 (chIFI35) contains an open reading frame (ORF) of 1,152 bp encoding a protein of 384 amino acids containing two conserved Nmi/IFI35 domain (NID) motifs. Tissue distribution analysis of chIFI35 in healthy and Newcastle disease (ND) virus-infected chickens indicated a positive correlation between chIFI35 mRNA transcription and ND viral loads in various tissues. The role of chIFI35 in regulation NDV replication were further assessed by up- or down-regulated chIFI35 expression in DF-1 cells transfected with plasmid harboring chIFI35, pCMV-3HA-chIFI35 or shRNA targeting chIFI35, pshRNA-chIFI35 plasmids. NDV replications in DF-1 cells were significantly reduced or slightly increased by over- or under-expression of the chIFI35 protein, respectively, indicating the role of chIFI35 in anti-NDV infection. Moreover, chIFI35 also involved in regulation of viral gene transcription and IFNs expression. The collected data were meaningful for research of chicken antiviral immunity and shed light on the pleiotropic antiviral effect of chIFI35 during NDV infection.

Subcutaneous angiolipoma in the scrotum: A case report.

BACKGROUND: Angiolipoma has been reported in many cases, and it often occurs in the skin of the trunk and limbs. However, angiolipoma in the scrotum is a rare disease with unknown etiology. This condition is difficult to diagnosis with other lumps in the scrotum. CASE SUMMARY: A 32-year-old man presented to the urinary department with a history of an enlarged left scrotum with increasing discomfort for about 5 years. Physical examination revealed that there were a palpable mass measuring about 7.0 cm × 6.5 cm in the left scrotum, with smooth surfaces but without tenderness or adhesion to the skin. Ultrasound showed that there was a hyperechoic mass under the skin of the top scrotum, about 72 mm × 64 mm × 21 mm in size, with clear borders, uneven internal echo, and abundant blood flow signals. Serum human chorionic gonadotropin and alpha-fetoprotein were in normal level. Subcutaneous mass resection at the bottom of the left scrotum was performed under local anesthesia with 1% lidocaine. Postoperative pathological examination resulted in a diagnosis of subcutaneous angiolipoma of the scrotum. No evidence of recurrence was found at 6 mo after surgery and there were no complaints of discomfort. CONCLUSION: Angiolipoma is an extremely rare type of benign tumor extremely rarely found in the scrotum, but needs to be considered when evaluating scrotal masses especially when the mass is solid. According to the characteristics of angiolipoma, surgical resection is the best treatment strategy and it is not prone to recurrence after resection.

Novel genotypes and multilocus genotypes of Enterocytozoon bieneusi in pigs in northwestern China: A public health concern.

Enterocytozoon bieneusi, the most important and common microsporidian species, inhabits in most animals and humans causing diarrhea and systemic diseases. The objectives of the present study were to examine the prevalence and genetic variability of E. bieneusi in pigs in Shaanxi province, northwestern China. A total of 560 pig faecal samples were collected from five different farms in Shaanxi province and molecularly characterized using multilocus genotyping (MLST) technology. High E. bieneusi infection rate (78.9%) was observed in these samples. 12 known and 22 possible novel ITS genotypes were identified, with the novel SZZD1 as the predominant genotype distributed in all age groups and pig farms. 32 (including 11 known and 21 novel ones) of them belong to the zoonotic group 1. MLST analysis showed that 109 ITS positive samples formed 87 distinct multilocus genotypes (MLGs). An incomplete linkage disequilibrium (LD) and clonal genetic structure of E. bieneusi were found in pigs in Shaanxi province. These findings indicated the complex population structures of E. bieneusi in pigs in Shaanxi province and provided baseline data for better understanding of the epidemiological status of E. bieneusi in this province.

Quantitative determination of the anticancer agent tubeimoside I in rat plasma by liquid chromatography coupled with mass spectrometry.

Tubeimoside I is an important component isolated from Bolbostemma paniculatum. Tubeimoside I has been demonstrated to possess many pharmacological activities, including anti-inflammatory, antitumor, and antitumor-promoting effects. The purpose of the present study was to examine in vivo pharmacokinetics and bioavailability of tubeimoside I in rats by using a liquid chromatography coupled with mass spectrometry quantitative detection method (LC/MS). The plasma samples were deproteinated, evaporated and reconstituted in 100 microl methanol prior to analysis. The separation was performed by Waters Symmetry C18 reversed-phase column (3.5 microm, 150 mm x 2.1mm, Waters Inc., USA) and a SB-C18 guard column (5 microm, 20 mm x 4.0mm). The mobile phase was a mixture of acetonitrile and water containing 5 microM NaAc (60:40, v/v). The method was validated within the concentration range 20-5000 ng/ml, and the calibration curves were linear with correlation coefficients >0.999. The lowest limit of quantitation (LLOQ) for tubeimoside I was 20 ng/ml in 0.1 ml rat plasma. The intra-assay accuracy and precision ranged from 92.4 to 104.9% and from 5.8 to 10.5%, respectively, while inter-assay accuracy and precision ranged from 94.2 to 95.0% and from 5.1 to 8.8%, respectively. The method was further applied to assess pharmacokinetics and oral bioavailability of tubeimoside I after intravenous and oral administration to rats. The oral bioavailability of tubeimoside I is only 0.23%, which indicates that tubeimoside I has poor absorption or undergoes acid-induced degradation. Practical utility of this new LC/MS method was confirmed in pilot pharmacokinetic studies in rats following both intravenous and oral administration.

Simultaneous determination of four active alkaloids from a traditional Chinese medicine Corydalis saxicola Bunting. (Yanhuanglian) in plasma and urine samples by LC-MS-MS.

A sensitive rapid method for the simultaneous determination of four major active alkaloids (dehydrocavidine, coptisine, dehydroapocavidine, and tetradehydroscoulerine, in abbreviation thereafter called YHL-I, YHL-II, YHL-III, and YHL-IV, respectively) from a Chinese traditional medicine Corydalis saxicola Bunting. (Yanhuanglian) in rat plasma and urine was established and validated. The assay for these substances in plasma and urine was based on HPLC coupled with tandem mass spectrometry (MS/MS) detection using multiple reaction monitoring mode (MRM) with berberine and clenbuterol as internal standards. The plasma and urine sample were deproteinated by adding methanol prior to liquid chromatography where separation was performed on a Luna column (5 microm, 100 x 2.00 mm) and an Agilent Zorbax SB-C18 guard column (5 microm, 20 x 4 mm). The method was validated with the concentration range 1-1000 ng/mL in plasma and 10-1000 ng/mL in urine for the four test compounds, and the calibration curves were linear with correlation coefficients >0.999. The lowest limits of quantitation for all four substances were 1 ng/mL in 0.1 mL rat plasma and 10 ng/mL in 0.1 mL urine. The intra-assay accuracy and precision in plasma ranged from 88.1 to 115.7% and 1.4 to 10.8%, respectively, while inter-assay accuracy and precision for YHL-I, YHL-II, YHL-III, and YHL-IV ranged from 96.2 to 113.2% and 0.4 to 16.9%, respectively. The intra-assay accuracy and precision for YHL-I, YHL-II, YHL-III, and YHL-IV in rat urine ranged from 96.1 to 112.9% and 1.2 to 8.3%, respectively, while inter-assay accuracy and precision ranged from 95.0 to 106.8% and 2.2 to 10.3%, respectively. The method was further applied to assess pharmacokinetics and urine excretion of the four alkaloids after oral and intravenous administration to rats. Practical utility of this new LC-MS-MS method was confirmed in pilot pharmacokinetic studies in rats following both intravenous and oral administration.

Bioavailabilty and pharmacokinetics of four active alkaloids of traditional Chinese medicine Yanhuanglian in rats following intravenous and oral administration.

Corydalis saxicola Bunting (Yanhuanglian) is an important component in various prescriptions in traditional Chinese medicine. Yanhuanglian has been demonstrated to possess many pharmacological activities, including antibacterial, antiviral and anticancer activities. The active fractions are dehydrocavidine, coptisine, dehydroapocavidine and tetradehydroscoulerine. The purpose of the present study was to examine in vivo pharmacokinetics and tissue distribution in rats by using high-performance liquid chromatography (HPLC) coupled with tandem mass spectrometry. Systemic clearance of the four active alkaloids in plasma was over 93% of hepatic blood flow, indicating they may be quickly eliminated via hepatic clearance. Less than 10% drugs was excreted via urine following intravenous and oral administration, suggesting that these four alkaloids may undergo significant metabolism in the body or the drug may be excreted via other routes other than urine. There was significantly lower excretion of these four alkaloids following oral than intravenous administration, suggesting a significant first pass effect after oral administration. There appeared to be wide distribution of those four alkaloids in rats as demonstrated by the higher apparent volume of distribution. Our results have also demonstrated that the four alkaloids can be absorbed following oral administration although there were less than 15% of drugs absorbed into systemic circulation. In summary, the favorable oral bioavailability properties of those four active alkaloids in rats make Yanhuanglian extract worth further investigation for improving oral bioavailability.

Outpatient procedures on the glans and anterior urethra under a new local anaesthesia: intracorpus spongiosum anaesthesia.

OBJECTIVE: To determine the efficacy of intracorpus spongiosum anaesthesia during minor procedures on the glans and anterior urethra in the outpatient clinic. METHODS: Sixty-nine consecutive male patients underwent various procedures on the glans penis or anterior urethra under intracorpus spongiosum anaesthesia, which was performed by injection of 3 mL of 1% lidocaine into the glans penis. The effect of this anaesthetic technique was assessed by questionnaire using a pain scale. RESULTS: Following injection of lidocaine, the anaesthetic effect was immediate and very satisfactory. The minor procedures, varying from 8 to 68 minutes in duration, were successfully completed under the new local anaesthesia. Of the 69 patients, 63 (91.3%) felt no pain and six (8.7%) reported either minor or moderate discomfort that was tolerable and could be ignored. There were no serious complications with this anaesthetic technique except for three patients (4.3%) who had instantaneous trance during lidocaine injection. CONCLUSION: Intracorpus spongiosum anaesthesia is an effective, simple, safe anaesthetic technique for minor procedures on the glans and anterior urethra in an outpatient setting. With this new local anaesthesia, the total cost of many procedures on the glans and anterior urethra can be markedly reduced.

Enhancement of palladium-porphyrin room temperature phosphorescence by alkaline earth metal in deoxycholate aggregates solution.

Room temperature phosphorescence (RTP) of three palladium (Pd)-phorphyrins in air-saturated solution of sodium deoxycholate (NaDC) aggregates was measured. RTP of Pd-meso-tetrakis (4-carboxyphenyl) porphyrin (Pd-TCPP) was obviously enhanced in NaDC-aggregates mediated by alkaline earth metal (AEM). Under the same experimental conditions, Ca(2+), Ba(2+) and Mg(2+) induced 200, 90 and 24 times greater enhancement in RTP of Pd-TCPP, respectively. It is ascribed to form the complex of NaDC-aggregate/AEM/Pd-TCPP in the system. The positively charged AEM has a strong capability of co-ordination with negatively charged carboxyl groups of NaDC and Pd-TCPP. The phosphor Pd-TCPP is confined in rigid NaDC-aggregates/AEM system by the coordination which decreases the probability of collision of phosphor with quenchers such as dissolved oxygen molecules and prolongs the lifetime of the phosphor on the triplet state. Long excited-state lifetimes resulted in great enhancement of Pd-TCPP phosphorescence. Observations by optical microscope showed that specific fan-like structures of NaDC were formed under the influence of AEM. Surface tension measurements supported a close interaction between Ca(2+) ions and anion aggregates of NaDC with 1:1 stoichiometric ratio. Due to its outstanding RTP behavior in NaDC-aggregates induced by Ca(2+), Pd-TCPP was used as a RTP probe to detect bovine serum albumin (BSA). A broad linear range from 1.0 × 10(-9) to 9.0 × 10(-7) g mL(-1) was obtained. Detection limit is 2.6 × 10(-11) g mL(-1), the relative standard deviation (n = 6) is 2.3% for 2.0 × 10(-9) g mL(-1) BSA.

[Effects of tourism disturbance on plant diversity in Qingshan Lake scenic area of Zhejiang Province].

From May 2007 to June 2008, an investigation was made on the changes of plant community in Qingshan Lake scenic area of Zhejiang Province under the effects of tourism disturbance. With the increase of tourism disturbance, the importance value of the plants was mainly fastened on a few species such as Pinus hwangshanensis, apt to decrease for tree and shrub species and to increase for herb species, and the individuals of the plants increased. The values of richness index (D) and diversity index (H) were in the order of medium disturbance > slight disturbance > severe disturbance, while the evenness index (J) value was in the order of medium disturbance > severe disturbance > slight disturbance. At the same vegetation layers, only a few species such as Cinnamomum camphora existed under different disturbances, and thereby, the similarity index values were smaller than 0.500. Slight disturbance affected coniferous forest most, with the average values of D, H, and J being the lowest (1.188, 1.056, and 0.697, respectively); severe disturbance affected broadleaf forest and shrub-herbage most, with the D value (2.013) of shrub-herbage and the H value (1.286) and J value (0.807) of broadleaf forest being the lowest; while medium disturbance was favorable to the increase of plant diversity and to the normal exertion of ecosystem function. The eco-safety of the structural elements of plant community in the scenic area was threatened to some extent, resulting in the reduction of indigenous species such as Sinocalycanthus chinensis and the incursion of exotic species as Setaria viridis.

Essential role for p53 and caspase-9 in DNA damaging drug-induced apoptosis in neuroblastoma IMR32 cells.

Neuroblastoma is a solid tumor of the sympathetic nervous system accounting for up to 10% of pediatric cancers and 15% of cancer-related deaths. It is a useful system for investigation of stress signal-mediated apoptosis as a tumor suppression mechanism. In this study, we present evidence that p53 mediates DNA damaging drug-induced apoptosis in IMR32 cells through the caspase-9 pathway. In summary, we define a molecular pathway for mediating DNA damaging drug-induced apoptosis in human neuroblastoma IMR32 cells and suggest that inactivation of essential components of this apoptotic pathway may confer drug resistance on neuroblastoma cells.

FSCB phosphorylation in mouse spermatozoa capacitation.

It is generally accepted that spermatozoa capacitation is associated with protein kinase A-mediated tyrosine phosphorylation. In our previous study, we identified the fibrous sheath CABYR binding protein (FSCB), which was phosphorylated by PKA. However, the phosphorylation status of FSCB protein during spermatozoa capacitation should be further investigated. To this aim, in this study, we found that phosphorylation of this 270-kDa protein occurred as early as 1 min after mouse spermatozoa capacitation, which increased over time and remained stable after 60 min. Immunoprecipitation assays demonstrated that the tyrosine and Ser/Thr phosphorylation of FSCB occurred during spermatozoa capacitation. The extent of phosphorylation and was closely associated with the PKA activity and spermatozoa motility characteristics. FSCB phosphorylation could be induced by PKA agonist DB-cAMP, but was blocked by PKA antagonist H-89.Therefore, FSCB contributes to spermatozoa capacitation in a tyrosine-phosphorylated format, which may help in further elucidating the molecular mechanism of spermatozoa capacitation.

[Summer habitat selection of alpine musk deer in Xinglongshan National Nature Reserve, Northwestern China].

During July and August from 2006 to 2008, summer habitat selection was studied in Alpine musk deer (Moschus sifanicus) in Xinglongshan National Natural Reserve in northwestern China. In total, seventy one musk deer utilized habitat plots and 246 random habitat plots were surveyed. Seventeen habitat characteristics were recorded and compared between the two habitat types, using Mann-Whitney U test and chi-square to compare the differences between utilized and random habitat plots, and principal component analysis (PCA) to determine the main factors influencing the habitat selection of musk deer. Mann-Whitney U test and chi-square was conducted to test whether there was significant difference between utilized and random plots. The results showed that musk deer prefers habitat with taller arbor height (7.57 +/- 0.83 ) m, higher food-plants abundance (12.97 +/- 1.80), increasing foliage cover for concealment, lower water dispersion, and higher anthropogenic disturbance. Furthermore, PCA results suggested that the arbor characteristics (arbor canopy and arbor DBH), altitude characteristic, food characteristics (ground-plant cover and food-plant abundance) and shrub characteristics (shrub canopy, shrub height and related arbor density) influence summer habitat selection patterns of alpine musk deer in Xinglongshan National Nature Reserve. The general pattern of summer habitat utilization and selection of alpine musk deer is an adaptive strategy to the changing food, concealment, water source and the physical condition of summer habitat.

[Prophylactic effect of inductive intravesical instillation of Bacillus Calmette-Guerin on postoperative recurrence of bladder cancer].

BACKGROUND & OBJECTIVE: Intravesical instillation is an important adjuvant therapy on preventing postoperative recurrence of superficial bladder transitional cell carcinoma, but the recurrence rate is still high. This study was to evaluate the prophylactic effect of intravesical instillation of hydroxycamptothecin (HYD) plus bacillus Calmette-Guerin (BCG) on postoperative recurrence of bladder transitional cell cancer. METHODS: A total of 45 bladder cancer patients who underwent TURBT or partial cystectomy were divided into two groups: 24 patients in combination group received single intravesical instillation of HYD in Week 1 after operation and regular intravesical instillation of BCG since Week 2; 21 patients in BCG group received regular intravesical instillation of BCG since Week 1 after operation. All the patients were followed up for 24 months. RESULTS: Three patients had recurrence at 2, 10, and 12 months after operation individually in BCG group; no recurrence developed in combination group. The recurrence rate was significantly higher in BCG group than in combination group (14.28% vs. 0, P<0.05). No serious adverse events and complication developed in both groups. CONCLUSION: Early use of single intravesical instillation of HYD plus subsequent regular intravesical instillation of BCG is markedly effective for preventing postoperative recurrence of bladder transitional cell cancer, with few adverse events.

Arterial anastomosis without sutures using ring pin stapler for clinical renal transplantation: comparison with suture anastomosis.

PURPOSE: The metal ring pin stapler was initially developed for microvascular surgery and there has been little experience of their use in larger vessels. We determined if the titanium ring pin coupling system could be safely and rapidly applied for arterial reconstruction in clinical renal transplantation. MATERIALS AND METHODS: The donor renal artery was end-to-end anastomosed to the internal iliac artery with titanium ring pin staplers in 36 patients. Anastomotic, clamp and total operative time and vascular problems were compared with those in the control group of sutured anastomosis in 39 transplant recipients. RESULTS: The completion of mechanical anastomosis required half the time of suture anastomosis. Mean clamp time was 17.8 minutes in the nonsuture group and 28.1 minutes in the control group. There was no significant difference in total operative time between the 2 groups. A small anastomotic line leak was noted in each group, which was temporary and self-limited. There were no postoperative anastomotic failures, postoperative bleeding episodes or need to revise the anastomosis (100% patency rate) in the 2 groups. Of the patients 61 were followed for 2 to 5 years. Transplant renal artery stenosis was observed in 1 patient in the nonsuture group but 3 in the control group. CONCLUSIONS: The anastomotic technique with the ring pin system is safe and simple, permitting an expeditious and everting anastomosis with a smooth intima-to-intima junction. Application of this technique may decrease warm ischemia time and the incidence of anastomotic artery stenosis, thus, improving outcomes.