[Source Profiles and Impact of Volatile Organic Compounds in Typical Industries in Luohe City].

Ten typical industries in Luohe City were selected for the sampling of organized emissions of volatile organic compounds （VOCs）, and 114 VOCs components of each sample were detected to analyze their source characteristics and effects. The results showed that VOCs emissions of packaging and printing were mainly composed of OVOC （60.9%）. In terms of the industrial coating, aromatic hydrocarbons （42.4%） and OVOC （38.9%） were the main VOCs species. The emissions of the footwear, furniture manufacturing, and paper industries were mainly composed of OVOC （32.3% - 42.6%） and aromatic hydrocarbons （20.7% - 33.7%）, with noticeable halogenated hydrocarbons. Chemical and pharmaceutical industries mainly emitted halogenated hydrocarbons, with the proportions of 59.3% and 46.6%, respectively. The emissions of the brick industry were primarily composed of alkane （62.7%）, and OVOC （48.5%）, and halogenated hydrocarbons （19.7%） were the main contributors to VOCs emissions of the thermal industry. OVOC （48.1%） and alkane （29.4%） were the dominant species for the food manufacturing industry. In the packaging and printing industry, acetone （14.8%）, isopropanol （14.0%）, ethylacetate （11.1%）, and toluene （10.2%） were the characteristic VOCs species. The emissions of industrial coating were dominated by isopropanol （25.6%）, toluene （15.0%）, m/p-xylene （12.4%）, and acetone （7.1%）. In the furniture manufacturing industry, m/p-xylene （15.8%）, followed by hexanal （15.1%）, 1,2-dichloroethane （9.6%）, and acetone （8.4%） were the characteristic VOCs species. The emissions of the footwear industry were dominated by acetone （18.9%）, toluene （18.1%）, methylene chloride （8.0%）, and acetaldehyde （6.8%）. The characteristic species of the chemical industry were methylene chloride （23.9%）, 1,2-dichloroethane （14.7%）, acetone （12.7%）, and trichloromethane （11.1%）, and those for the pharmaceutical industry were bromoethane （36.7%）, acetone （19.2%）, benzene （5.0%）, and vinyl acetate （3.0%）. The emissions of the brick industry were mainly ethane, propane, ethylene, and benzene. Acetone, toluene, acetylene, and acetaldehyde were the primary VOCs species in the paper industry. The emissions of the food manufacturing industry were dominated by acetaldehyde, n-pentane, acrolein, and n-heptane. The emissions of the thermal industry were characterized by acetone, acetaldehyde , benzene, and toluene. Although different industries emitted various characteristic VOCs species, in general, acetone, isopropanol, benzene, toluene, m/p-xylene, ethane, acetaldehyde, and methylene chloride were the main characteristic species in most industries in Luohe. OVOC and aromatic hydrocarbons had higher contributions to ozone generation potential （OFP）, and aromatic hydrocarbons contributed over 80.0% to secondary organic aerosol formation potential （SOAP）. The source reactivity of ozone [SR（O3）] of the food and furniture manufacturing industries were higher, with values of 3.7 g·g-1 and 3.5 g·g-1, respectively, whereas the source reactivity of secondary organic aerosol SR（SOA） of the industrial coating, furniture manufacturing, and footwear industries were higher, with the values of 0.021, 0.017, and 0.014 g·g-1. Hence, the food manufacturing, industrial coating, and furniture manufacturing industries should be the primary industries for the collaborative control of PM2.5 and ozone in Luohe City, of which the furniture manufacturing industry was the top priority.

Impact of smart physical education assignment on physical health of male college students / 中国学校卫生

Objective@#To explore the impact of smart physical education assignment on physical health of male university students, so as to provide theoretical support and practical references for physical health improvement of male university students and implementing smart sports assignments.@*Methods@#From September 2023 to January 2024, 317 sophomore male students from six Taekwondo elective classes at Hunan Institute of Engineering were selected and were randomly divided into an intervention group (n=157) and a control group (n=160). The intervention group was given sports assignments twice a week through smart means with an intervention duration of 15 weeks, each time for 25-35 minutes, in addition to the teaching according to the public course syllabus, while the control group was taught according to the public course syllabus. The physical and health indicators of both groups were tested before and after intervention,then the differences in various physical health indicators between two groups of students before and after intervention were compared through ttest and Mann-Whitney U test.@*Results@#After the intervention, the vital capacity, 50 m run, sitandreach, 1 000 m run, and pullup scores of the intervention group significantly improved compared to those before intervention. The scores improved from (3 918.27±737.34)mL, 7.88(7.53,8.45)s, 9.80(2.70,15.75) cm, 4.30(4.12,4.50) min and 3.00(0.00,7.50) times to (4 574.19±800.61) mL, 7.65(7.37,8.12)s, 17.20(11.80,21.55)cm, 4.13(3.58,4.31)min and 5.00(1.00,10.00) times,respectively (t/Z=-7.60, 2.61, -8.39, 5.62, -2.72, P<0.05). Before intervention, there was no statistically significant difference in physical health indicators between the intervention group and the control group (P>0.05).After intervention,the scores of the intervention group on the vital capacity,50 m run,sitandreach,1 000 m run and pullup, were significantly higher than those of the control group [(4 310.97±808.90)mL, 7.75(7.40,8.30)s, 14.10(8.42,17.87)cm, 4.29(4.08,4.45)min and 4.00(1.00,7.00) times] (t/Z=2.91, -4.55, -4.75, -4.15, 2.58, P<0.05).@*Conclusions@#Having 25-35 min smart physical education assignment twice a week can effectively improve physical health level of male college students. It is recommended to assign appropriate amount of smart sports homework to improve physical health level of college students, while ensuring the amount and intensity of physical activity in public physical education courses.

Clinical and chest computed tomography features associated with severe Chlamydia psittaci pneumonia diagnosed by metagenomic next-generation sequencing: A multicenter, retrospective, observational study.

Chlamydia psittaci pneumonia is a rare disease with varying clinical presentations. Here, we aimed to investigate the clinical and chest computed tomography (CT) features of severe psittacosis pneumonia. Clinical data of 35 patients diagnosed with psittacosis pneumonia were retrospectively analyzed using metagenomic next-generation sequencing. The patients were classified into severe (n = 20) and non-severe (n = 15) groups. The median age of patients was 54 years, and 27 patients (77.1%) had a definite history of bird contact. Severe patients had more underlying comorbidities and were more prone to dyspnea and consciousness disorders than non-severe patients. The neutrophil count and D-dimer, lactate dehydrogenase, interleukin (IL)-2, IL-6, and IL-10 levels were higher, whereas the lymphocyte, CD3 + T cell, and CD4 + T cell counts, CD4+/CD8 + T cell ratio, and albumin level were substantially lower in severe patients than in non-severe patients. Chest CT findings of severe patients revealed large areas of pulmonary consolidation, and ground-glass opacities were observed in some patients, with a higher risk of involving multiple lobes of the lungs and pleural effusion. One patient died of multiple organ failure, whereas the condition of the other 34 patients improved, and they were discharged from the hospital. Patients with severe psittacosis pneumonia often have underlying comorbidities and are prone to developing dyspnea, consciousness disorder, and lesions in both lungs. Serum D-dimer, IL-2, IL-6, and IL-10 levels and lymphocyte, CD3 + T cell, and CD4 + T cell counts are associated with disease severity.

Chimeric Mouse Generation by ES Cell Blastocyst Microinjection and Uterine Transfer.

The ability to generate chimeric mice through microinjecting embryonic stem (ES) cells into blastocysts is a critical step for the conventional ES cell-mediated knockout technology. In recent years, designer nuclease-based methods, especially the CRISPR technology, have substantially decreased the needs for blastocyst microinjection. However, this method has still remained as a valuable technique for generating sophisticated genetic models as well as for stem cell research. In this chapter, we describe the detailed procedures used in our laboratory on how to use ES cells to produce chimeric mice, including derivation and inactivation of MEF feeder cells, culturing and handling of mouse ES cells, collection and microinjection of blastocysts, and finally implantation of injected blastocysts into the uteri of pseudopregnant surrogate mothers.

Using CRISPR/Cas9 for Gene Knockout in Immunodeficient NSG Mice.

NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG) mice are an immunodeficient strain that enables human cell xenografts. However, NSG mice possess a complex genetic background that would complicate cross-breeding with other inbred transgenic or knockout mouse strains to establish a congenic strain with a desired genetic modification in the NSG background. Newly developed clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 technology enables modification of the mouse genome at the zygote stage without the need for extensive cross-breeding or the use of embryonic stem cells. In this chapter, we use the knockout of the X-linked Cybb gene as an example to describe our procedures for genetically modifying NSG mice using the CRISPR/Cas9 method. Briefly, two sgRNAs were designed and made to target exon 1 and exon 3 of the Cybb gene, and either sgRNA was then microinjected together with Cas9 mRNA into fertilized eggs collected from NSG mice. The injected embryos are subsequently transferred into the oviducts of pseudopregnant surrogate mothers. Offspring born to the foster mothers were genotyped by PCR and DNA sequencing. In this chapter, we describe our experiment procedures in detail and report our genotyping results for demonstrating that NSG mice can be genetically modified using the CRISPR/Cas9 technology in a highly efficient manner.

Generation of Conditional Knockout Mice by Sequential Insertion of Two loxP Sites In Cis Using CRISPR/Cas9 and Single-Stranded DNA Oligonucleotides.

Conditional knockout (cKO) mice are extremely valuable for biomedical research because they enable detailed analyses of gene functions in a tissue- or temporally-specific fashion. The conventional method for generating cKO mice is time consuming and labor intensive, which involves making a large gene-targeting construct, transfecting and screening many embryonic stem (ES) cell clones, injecting positive ES clones into blastocysts to produce chimeric mice, and breeding the chimeras to transmit the targeted gene through the germline. Recently developed CRISPR technology has revolutionized the way genetically engineered organisms are created. Knockout and knockin mice can now be made by directly injecting zygotes with Cas9, sgRNA, and donor DNA. In theory, cKO mice can be generated by simultaneously inserting two loxP sites using two sgRNAs and two oligonucleotides as donors, but in practice the probability of obtaining cKO mice in one step is still very low, partly because the efficiency of oligo-mediated knockin is much lower than non-homologous end joining (NHEJ) and partly because co-cutting juxtaposed sites in one allele at the same time often leads to the deletion of the entire fragment between the two cutting sites. Therefore, many laboratories prefer to insert the two loxP sites sequentially, i.e., generating mice with one loxP first and then use embryos collected from these mice to insert the second loxP site. In this chapter, we describe our procedures and timeline using this sequential method to make a Six6 cKO mouse line as a demonstration of its feasibility.

Cutting Edge: ß-Catenin-Interacting Tcf1 Isoforms Are Essential for Thymocyte Survival but Dispensable for Thymic Maturation Transitions.

T cell factor 1 (Tcf1) is essential for T cell development; however, it remains controversial whether ß-catenin, a known coactivator of Tcf1, has a role. Tcf1 is expressed in multiple isoforms in T lineage cells, with the long isoforms interacting with ß-catenin through an N-terminal domain. In this study, we specifically ablated Tcf1 long isoforms in mice (p45-/-mice) to abrogate ß-catenin interaction. Although thymic cellularity was diminished in p45-/- mice, transition of thymocytes through the maturation stages was unaffected, with no overt signs of developmental blocks. p45-/- thymocytes showed increased apoptosis and alterations in transcriptome, but these changes were substantially more modest than in thymocytes lacking all Tcf1 isoforms. These data indicate that Tcf1-ß-catenin interaction is necessary for promoting thymocyte survival to maintain thymic output. Rather than being dominant-negative regulators, Tcf1 short isoforms are adequate in supporting developing thymocytes to traverse through maturation steps and in regulating the expression of most Tcf1 target genes.

Heparin Promotes Cardiac Differentiation of Human Pluripotent Stem Cells in Chemically Defined Albumin-Free Medium, Enabling Consistent Manufacture of Cardiomyocytes.

Cardiomyocytes can be differentiated from human pluripotent stem cells (hPSCs) in defined conditions, but efficient and consistent cardiomyocyte differentiation often requires expensive reagents such as B27 supplement or recombinant albumin. Using a chemically defined albumin-free (E8 basal) medium, we identified heparin as a novel factor that significantly promotes cardiomyocyte differentiation efficiency, and developed an efficient method to differentiate hPSCs into cardiomyocytes. The treatment with heparin helped cardiomyocyte differentiation consistently reach at least 80% purity (up to 95%) from more than 10 different hPSC lines in chemically defined Dulbecco's modified Eagle's medium/F-12-based medium on either Matrigel or defined matrices like vitronectin and Synthemax. One of heparin's main functions was to act as a Wnt modulator that helped promote robust and consistent cardiomyocyte production. Our study provides an efficient, reliable, and cost-effective method for cardiomyocyte derivation from hPSCs that can be used for potential large-scale drug screening, disease modeling, and future cellular therapies. Stem Cells Translational Medicine 2017;6:527-538.

Induced pluripotent stem cell transplantation in the treatment of porcine chronic myocardial ischemia.

BACKGROUND: This study was designed to test the effects of induced pluripotent stem cell (iPSC) in the treatment of chronic myocardial ischemia. METHODS: The reprogramming of passage 3 myocardial fibroblasts was performed by using the lentiviral vector containing 4 human factors: OCT4, SOX2, KLF4, and c-MYC. The iPSC colonies at P12-17 were allogeneically transplanted into ischemic myocardium of 10 swine by direct injection. Cohorts of 2 animals were sacrificed at 2, 4, 6, 8, and 12 weeks after injection. RESULTS: No signs of graft versus host disease were evident at any time points. At 2 weeks, clusters of SSEA-4-positive iPSCs were detected in the injected area. At 4 to 8 weeks, these cells started to proliferate into small spheres surrounded by thin capsules. At 12 weeks the cell clusters still existed, but decreased in size and numbers. The cells inside these masses were homogeneous with no sign of differentiation into any specific lineage. Increased smooth muscle actin or vWF positive cells were found inside and around the iPSC clusters, compared with non-injected areas. By real-time polymerase chain reaction, the levels of VEGF, basic FGF, and ANRT expression were significantly higher in the iPSC-treated myocardium compared with untreated areas. These results suggest that iPSCs contributed to angiogenesis. CONCLUSIONS: Allogeneically transplanted pig iPSCs proliferated despite an ischemic environment in the first 2 months and survived for at least 3 months in immunocompetent hosts. Transplanted iPSCs were also proangiogenic and thus might have beneficial effects on the ischemic heart diseases.

Purification of plasmid and BAC transgenic DNA constructs.

Pronuclear microinjection is the most used method for generating transgenic mice. The quality of DNA to be microinjected is a key determinant of the success rate of this method. DNA purity is a critical factor because trace amounts of many substances, when microinjected into the pronucleus of the fertilized egg, can kill or prevent the further development of the embryo. Avoiding all contaminants is not a trivial issue, because most transgenic fragments need to be purified from agarose gels. Small particles and viscous materials in the DNA solution can also dramatically reduce the efficiency of microinjection because they tend to clog the injection needles. DNA shearing or breakage during purification and microinjection is also a potential problem, particularly when linearized bacterial artificial chromosomes (BAC) DNAs are used. The overall quantity and the final DNA concentration are also important considerations, because egg -pronuclei are very sensitive to the amount of foreign DNA. In this chapter, we first discuss the general guidelines and cautions for preparing microinjection-quality DNA, and then describe in detail two -protocols, one for gel purification of transgenic fragments from plasmid vectors and the other for isolating high-quality BAC DNA from bacteria.