Production of Transgenic Pigs Mediated by Pseudotyped Lentivirus and Sperm

Sperm-mediated gene transfer can be a very efficient method to produce transgenic pigs, however, the results from different laboratories had not been widely repeated. Genomic integration of transgene by injection of pseudotyped lentivirus to the perivitelline space has been proved to be a reliable route to generate transgenic animals. To test whether transgene in the lentivirus can be delivered by sperm, we studied incubation of pseudotyped lentiviruses and sperm before insemination. After incubation with pig spermatozoa, 62±3 lentiviral particles were detected per 100 sperm cells using quantitative real-time RT-PCR. The association of lentivirus with sperm was further confirmed by electron microscopy. The sperm incubated with lentiviral particles were artificially inseminated into pigs. Of the 59 piglets born from inseminated 5 sows, 6 piglets (10.17%) carried the transgene based on the PCR identification. Foreign gene and EGFP was successfully detected in ear tissue biopsies from two PCR-positive pigs, revealed via in situ hybridization and immunohistochemistry. Offspring of one PCR-positive boar with normal sows showed PCR-positive. Two PCR-positive founders and offsprings of PCR-positive boar were further identified by Southern-blot analysis, out of which the two founders and two offsprings were positive in Southern blotting, strongly indicating integration of foreign gene into genome. The results indicate that incubation of sperm with pseudotyped lentiviruses can incorporated with sperm-mediated gene transfer to produce transgenic pigs with improved efficiency

Experiments on a New Material for the Ecological Protection of Rock Slopes

AbstractFrom 2000, people use the ecological protection to reinforce rock slopes. These ecological protection methods not only can reduce the usage of concrete, but also can restore the degraded ecosystem of slopes. With the rapid development of infrastructures, many materials for ecological protection have been created. However, there are many defects in these materials, such as expensive price, weak strength in early period, easily slide or been eroded when encountered with rainfall. These defects are the problems awaiting solution in the development of slope ecological protection. In this paper, the author attempts to use cement as adhesive in a new material for the ecological protection, and carry out laboratory tests and field tests to verify the feasibility and reliability of this material. Based on existing research and the testing results, the author proposed the prescription of this new material, which can satisfy the survival requirement of vegetation on the slopes, lower costs and enhance the strength in early period of ecological protection as well

Tuina combined with moxibustion for 28 cases of lung-wei insecurity-type sweating syndrome in children

目的:探讨推拿配合艾灸治疗肺卫不固型小儿汗症的临床疗效。方法:采用小儿推拿结合艾灸治疗28例肺卫不固型小儿汗症患儿,推拿隔日一次,艾灸隔三日一次; ,治疗4周后观察临床疗效。结果:推拿配合艾灸治疗肺卫不固型小儿汗证的总有效率为92.85%。结论:推拿配合艾灸治疗肺卫不固型小儿汗证疗效肯定,值; 得临床上推广应用。Objective To explore the clinical efficacy of tuina combined with; moxibustion for treatment of lung-wei insecurity-type sweating syndrome; in children. Methods Infantile tuina combined with moxibustion was; conducted for 28 cases of lung-wei insecurity-type sweating syndrome in; children. Tuina was carried out for once every other day, and; moxibustion was conducted for once every other three days. Clinical; efficacy was observed after treatment for 4 weeks. Results The total eff; ective rate of tuina combined with moxibustion for treatment of lung-wei; insecurity-type sweating syndrome in children was 92.85%. Conclusion The; efficacy of tuina combined with moxibustion for treatment of lung-wei; insecurity-type sweating syndrome in children is definite, which is; worthy of being popularized clinically.Supported by Inheritance venation and academic idea study of Huxiang; five meridians combination acupuncturemoxibustion and tuina academic; schoo

Integrated Analysis of a Ferroptosis-Related LncRNA Signature for Evaluating the Prognosis of Patients with Colorectal Cancer

LncRNAs have been well known for their multiple functions in the tumorigenesis, development, and relapse of colorectal cancer (CRC). Accumulating studies demonstrated that the expression of lncRNAs can be regulated by ferroptosis, a biological process that has been revealed to suppress CRC progression. However, the functions and clinical implications of ferroptosis-associated lncRNAs in CRC remain largely unknown. We, herein, aim to construct a prognostic signature with ferroptosis-related lncRNAs for the prognostic estimation of CRC patients. Firstly, we identified the lncRNAs related to ferroptosis based on the RNA-Seq data of CRC from the TCGA database. The univariate and multivariate Cox analyses were then performed to establish a prognostic signature composed of eight ferroptosis-related lncRNAs (AL161729.4, AC010973.2, CCDC144NL-AS1, AC009549.1, LINC01857, AP003555.1, AC099850.3, and AC008494.3). Furthermore, we divided the CRC patients into high- and low-risk groups based on the signature and found the overall survival (OS) of patients in the high-risk group was significantly shorter than that in the low-risk group (p = 3.31 × 10−11). Moreover, the patients in the high-risk groups had shorter recurrence-free survival (RFS) (p = 6.5 × 10−3) and disease-free survival (DFS) (p = 4.27 × 10−4), as well as higher tumor recurrence rate. Additionally, we found that the oncogenic pathways were enriched in the high-risk group, whereas the ferroptosis pathway that probably repressed CRC development was enriched in the low-risk group. In summary, our signature may provide a theoretical foundation for not only accurate judgment for prognosis but also evaluation for recurrence and metastasis in CRC patients

Transportation characteristics of δ 13C in the plants-soil-bedrock-cave system in Chongqing karst area

Here we use an analytical method to determine δ 13C in local plants and organic matter in the soils above Furong cave, Chongqing, China. We also monitored δ 13C in dissolved inorganic carbon (DIC) of drip water, δ 13C of active deposits under the drip waters, and the concentration of air CO2 (PCO2). Based on these, we preliminarily studied the transportation characteristics of stable carbon isotope (13C) in cave system of the subtropical karst area. The average δ 13C value of 27 local plant samples, which belong to 16 families, was −32‰ and the weighted δ 13C for surface dry biomass was −33‰. We found that for 54 soil samples collected from 5 soil profiles, δ 13C of soil organic matters was −22‰, which could be attributed to the different transportation rates of stable carbon isotopes during the decomposition of plants and organic matters in soils. The relatively lighter 12C tended to transfer into gaseous CO2, which made the relatively heavier 13C concentrated in the soils. On the basis of monitoring of DIC-δ 13C in drip waters from July 2009 to June 2010, we found that values in winter months were heavier and values in summer months were lighter in general, the reason of which was that in summer months, both the temperature and the humidity were comparatively higher, resulted in more CO2 with lighter δ 13C generated from organic matters decomposition and plants respiration. The average DIC-δ 13C value was −11‰, about 11‰ heavier than the δ 13C of organic matters in soils, which proved that part of DIC in cave drip water was sourced from dissolution of inorganic carbonate (host rock, with heavier δ 13C). As for the δ 13C of active deposits at five drip water sites in Furong cave, they had almost the same variation with relatively light values. In other words, these active speleothems were deposited at equilibrium conditions for isotopic fractionation. These results suggest that the carbon isotopic information of speleothems could be used to track the evolution of local vegetation in certain situations

Transportation characteristics of δ 13C in the plants-soil-bedrock-cave system in Chongqing karst area

Here we use an analytical method to determine δ 13C in local plants and organic matter in the soils above Furong cave, Chongqing, China. We also monitored δ 13C in dissolved inorganic carbon (DIC) of drip water, δ 13C of active deposits under the drip waters, and the concentration of air CO2 (PCO2). Based on these, we preliminarily studied the transportation characteristics of stable carbon isotope (13C) in cave system of the subtropical karst area. The average δ 13C value of 27 local plant samples, which belong to 16 families, was −32‰ and the weighted δ 13C for surface dry biomass was −33‰. We found that for 54 soil samples collected from 5 soil profiles, δ 13C of soil organic matters was −22‰, which could be attributed to the different transportation rates of stable carbon isotopes during the decomposition of plants and organic matters in soils. The relatively lighter 12C tended to transfer into gaseous CO2, which made the relatively heavier 13C concentrated in the soils. On the basis of monitoring of DIC-δ 13C in drip waters from July 2009 to June 2010, we found that values in winter months were heavier and values in summer months were lighter in general, the reason of which was that in summer months, both the temperature and the humidity were comparatively higher, resulted in more CO2 with lighter δ 13C generated from organic matters decomposition and plants respiration. The average DIC-δ 13C value was −11‰, about 11‰ heavier than the δ 13C of organic matters in soils, which proved that part of DIC in cave drip water was sourced from dissolution of inorganic carbonate (host rock, with heavier δ 13C). As for the δ 13C of active deposits at five drip water sites in Furong cave, they had almost the same variation with relatively light values. In other words, these active speleothems were deposited at equilibrium conditions for isotopic fractionation. These results suggest that the carbon isotopic information of speleothems could be used to track the evolution of local vegetation in certain situations

MOESM1 of Dynamic and specific immune responses against multiple tumor antigens were elicited in patients with hepatocellular carcinoma after cell-based immunotherapy

Additional file 1. Additional figures and tables

Map of lentiviral vector.

<p>Map of lentiviral vector.</p