Genomic Dissection through Whole-Genome Resequencing of Five Local Pig Breeds from Shanghai, China.

China has rich genetic resources of local pig breeds. In this study, whole-genome resequencing was performed on five Shanghai local pig breeds, aiming to analyze their population genetic structure and unique genomic characteristics. Tens of millions of single nucleotide variants were obtained through the resequencing of a total of 150 individual pigs from five local pig breeds (Meishan, Fengjing, Shawutou, Pudong White, and Shanghai White) after mapping them with the pig reference genome of Sus scrofa 11.1. The results of admixture structure analysis also clearly demonstrated the genetic differences between the Shanghai local pig breeds and the three commercial pig breeds (Duroc, Landrace, and Yorkshire). The genetic infiltration of Landrace and Yorkshire pig breeds in the SHW breed was detected, which is consistent with the early history of crossbreeding in this breed. Selective sweep analysis between four indigenous Shanghai pig breed populations and three commercial pig breed populations identified 270 and 224 genes with selective signatures in the commercial and indigenous Shanghai pig populations, respectively. Six genes (TGS1, PLAG1, CHCHD7, LCORL, TMEM68, and TMEM8B) were found to be associated with animal growth in the commercial pig population through gene enrichment and protein-protein interaction analysis. In contrast, the MSRB3 gene in the indigenous Shanghai pig population was significantly under selection, which correlated with the long pendulous ear phenotype of the indigenous Shanghai pig population. In conclusion, this study is the first genomic profiling of five representative local pig breeds in Shanghai, which provides molecular genetic data and foundations for better conservation and utilization of local pig breed resources in Shanghai, China.

Preliminary Results on Preparation and Performance of a Self-Emulsifying Waterborne Epoxy Curing Agent at Room Temperature.

Polyethylene glycol 1000 (PEG1000) and epoxy resin E20 were used to synthesize the E20/PEG1000 polymer (EP1K), which was later transformed into a self-emulsifying water-based epoxy curing agent by reacting with m-Xylylenediamine (MXDA). The effects of molecular weight, the molar ratio of the raw materials, the catalyst dosage, and the different co-solvents on the properties of the prepared curing agent were systematically explored. The infrared absorption spectra of E20, EP1K, and the water-based epoxy curing agent were compared and analyzed. The coating properties of the waterborne epoxy varnish, which was based on water-based epoxy curing agents to emulsify and cure the resin E44, were systematically tested. The results demonstrated that with a molar ratio of 1:1:4 of PEG1000, E20, and MXDA, the boron trifluoride etherate (BF3·Et2O) as catalyst accounts for 0.3% of the total mass of E20 and PEG1000, and an applicable period of 3 h for the prepared varnish, the anti-corrosion performance, and mechanical properties of the coatings were excellent.

Nephrotoxicity of immune checkpoint inhibitor combination therapy in patients with advanced renal cell carcinoma: a meta-analysis.

PURPOSE: Few data are available regarding the nephrotoxicity of immune checkpoint inhibitor (ICI) combination therapy in advanced renal cell carcinoma (RCC). This study aimed to investigate the nephrotoxicity of ICI-based combination therapy versus standard of care sunitinib in patients with advanced RCC. METHODS: We searched Embase/PubMed/Cochrane Library for relevant randomized controlled trials (RCTs). Treatment-related nephrotoxicities including increase of creatinine and proteinuria were analyzed by Review Manager 5.4 software. RESULTS: Seven RCTs involving 5239 patients were included. The analysis showed that ICI combination therapy had similar risks of any grade (RR = 1.03, 95% CI: 0.77-1.37, P = 0.87) and grade 3-5 (RR = 1.48, 95% CI: 0.19-11.66, P = 0.71) increased creatinine compared with sunitinib monotherapy. However, ICI combination therapy was associated with significantly higher risks of any grade (RR = 2.33, 95% CI: 1.54-3.51, P < 0.0001) and grade 3-5 proteinuria (RR = 2.25, 95% CI: 1.21-4.17, P = 0.01). CONCLUSIONS: This meta-analysis suggests that ICI combination therapy shows more nephrotoxicity of proteinuria than sunitinib in advanced RCC, which deserves a high attention in the clinic.

Identification of tRNA-derived small RNAs and their potential roles in porcine skeletal muscle with intrauterine growth restriction.

Intrauterine growth restriction (IUGR) in humans often manifests as poor growth and delayed intellectual development, whereas in domestic animals it results in increased mortality. As a novel epigenetic regulatory molecule, tRNA-derived small RNAs (tsRNAs) have been reported to be involved in many biological processes. In this study, pigs (35d) were used as a model to characterize tsRNAs by sequencing in normal and IUGR porcine skeletal muscle. A total of 586 tsRNAs were identified, of which 103 were specifically expressed in normal-size pigs and 38 were specifically expressed in IUGR pigs. The tsRNAs formed by splicing before the 5' end anti codon of mature tRNA (tRF-5c) accounted for over 90% of tsRNAs, which were significantly enriched in IUGR pigs than in normal-size pigs. Enriched pathways of differentially expressed tsRNAs target genes mainly included metabolic pathways, Rap1 signaling pathway, endocytosis, mTOR signaling pathway, and AMPK signaling pathway. Regulatory network analysis of target genes revealed that IGF1 was one of the most important molecules of regulatory nodes in IUGR and normal porcine skeletal muscle. In addition, IGF1 was found to be one of the target genes of tRF-Glu-TTC-047, which is a highly expressed tsRNA in IUGR pigs. The findings described herein uncover the role of tsRNAs in IUGR porcine skeletal muscle development, thus providing insights into the prevention and treatment of IUGR in mammals.

MiR-29a Family as a Key Regulator of Skeletal Muscle Dysplasia in a Porcine Model of Intrauterine Growth Retardation.

MicroRNAs (miRNAs) play an essential role in many biological processes. In this study, miRNAs in the skeletal muscle of normal and intrauterine growth retardation (IUGR) neonatal piglets were identified by sequencing, and canonical miRNAs were functionally validated in vitro. A total of 403 miRNAs were identified in neonatal piglet skeletal muscle, among them 30 and 46 miRNAs were upregulated and downregulated in IUGR pigs, respectively. Upregulated miRNAs were mainly enriched in propanoate metabolism, endocytosis, beta-Alanine metabolism, gap junction, and tumor necrosis factor signaling pathway. Down-regulated miRNAs were mainly enriched in chemical carcinogenesis-receptor activation, endocytosis, MAPK signaling pathway, insulin resistance, and EGFR tyrosine kinase inhibitor resistance. Co-expression network analysis of umbilical cord blood and skeletal muscle miRNAs showed that the miR-29 family is an essential regulator of IUGR pigs. The dual-luciferase reporter system showed that IGF1 and CCND1 were target genes of the miR-29 family. Transfection of IUGR pig umbilical cord blood exosomes and miR-29a mimic significantly inhibited cell proliferation and promoted the expression of cellular protein degradation marker genes Fbxo32 and Trim63. In summary, these results enrich the regulatory network of miRNAs involved in skeletal muscle development in IUGR animals.

Three-Dimensional Cuprous Iodide Framework with Intrinsic Broadband Red-to-Near-Infrared Light Emission.

Herein, a new organic-inorganic hybrid cuprous iodide of [(Me)2-DABCO]Cu6I8 was prepared and structurally characterized with a novel three-dimensional (3D) [Cu6I8]2- framework. Significantly, this 3D cuprous iodide displays infrequent broadband red-to-near-infrared light emission (600-1000 nm) stemming from the radiative recombination of self-trapped excitons.

Chemical composition, energy content, and amino acid digestibility in Cyperus esculentus co-products fed to growing pigs.

This study was conducted to determine the chemical composition, DE, ME, and NE contents, and the apparent and standardized ileal digestibility (AID and SID) of AA in Cyperus esculentus co-products fed to growing pigs. The five C. esculentus co-products included expeller-pressed C. esculentus cake (EPCC), cold-pressed C. esculentus cake (CPCC), solvent-extracted C. esculentus meal (SECM), C. esculentus distillers's dried grains with solubles (CE DDGS), and C. esculentus meal (CEM). In Exp. 1, a total of 36 crossbred growing pigs (Duroc × Landrace × Yorkshire; BW: 50.12 ± 2.91 kg) were fed one of six diets in a completely randomized design. The diets included a corn-soybean meal basal diet and five experimental diets containing 24.31% C. esculentus co-products. In Exp. 2, 12 same breed of growing pigs (BW: 47.12 ± 3.2 kg), surgically fitted with a T-cannula in the distal ileum, were allotted to one of four experimental diets in a 2-period Youden Square design. The diets included one N-free diet and three experimental diets containing 50% C. esculentus co-products (including EPCC, SECM, and CE DDGS). Results indicated that the SECM and CE DDGS had the greatest contents of starch and CP, respectively. The contents of CF, NDF, and ADF were the greatest in CEM and the lowest in SECM. On a DM basis, the DE, ME, predicted NE, and apparent total tract digestibility (ATTD) of GE values of the 5 C. esculentus co-products ranged from 1,203 to 3,897 kcal/kg, 1,127 to 3,621 kcal/kg, 536 to 2,871 kcal/kg, and 28% to 79%, respectively. The EPCC and CPCC had the greatest DE, ME, and predicted NE values, and CPCC, EPCC, and SECM had the greatest ATTD of GE, whereas CEM had the lowest DE, ME, NE, and ATTD of GE (P < 0.001). The NDF and ADF were negatively correlated with DE, ME, and NE (P < 0.05). The AID and SID of CP varied from 53.57 % to 57.86% and from 69.99% to 87.85%, respectively. The EPCC and SECM had greater SID of CP, Ile, Met, Val, Asp, Cys, and Tyr compared to those of CE DDGS (P < 0.05). These results indicated that the chemical composition, DE, ME, and NE as well as the most AA digestibility of C. esculentus co-products obtained from different processing techniques varied greatly. Based on the energy contents and AA digestibility, the EPCC is a better feedstuff for growing pigs compared with the other 4 C. esculentus co-products.

Expression Characteristics of microRNA in Pig Umbilical Venous Blood and Umbilical Arterial Blood.

As the medium of material exchange between mother and fetus, umbilical cord blood is closely connected with fetal development. microRNA (miRNA) has a wide range of biological functions and has high flow characteristics. Small RNA sequencing of pig umbilical venous blood (UVB) and umbilical arterial blood (UAB) revealed that a total of 302 miRNAs were identified, and 106 and 22 miRNAs were specifically expressed in the UVB and UAB, respectively. Using the two methods of differential expression multiple and differential expression percentage, it is found that only 35% of the highly expressed miRNAs in the UVB by the two analysis modes overlap, but 56.25% of the enriched signal pathways are the same. Only 20% of the highly expressed miRNAs in the UAB overlap, but 62.07% of the signal pathways are the same. Further analysis revealed that miR-423 can be used as a characteristic miRNA of UVB and has the potential to treat muscle-related diseases. miR-122-5p can be used as a characteristic miRNA of UAB and may help to improve liver- and brain-related diseases. In summary, these results enrich understanding of miRNA in mother-fetal communication and provide a reference for the development and application of porcine cord blood products.

Enhancement of the photoluminescence efficiency of hybrid manganese halides through rational structural design.

Five new zero-dimensional hybrid manganese halides based on discrete [MnCl4]2- tetrahedrons were prepared and used as highly efficient green-light emitters. Through rational management of organic cations to tailor the MnMn separation distances between neighboring [MnCl4]2- tetrahedrons, the photoluminescence quantum yield increased significantly from 7.98% to 81.11%.

Design and synthesis of sinomenine isoxazole derivatives via 1,3-dipolar cycloaddition reaction.

A novel structure of sinomenine isoxazole derivatives is synthesised from sinomenine hydrochloride and aromatic aldehydes and requires six steps. 19 target compounds have been obtained in good yields. The sinomenine hydrochloride transforms to 4-alkynyl sinomenine, which is a key intermediate product to synthesise the target sinomenine isoxazole compounds, after a neutralisation reaction with ammonia and substitution reaction with 3-chloropropyne. Another key intermediate product is 1,3-dipole, which can be obtained from aromatic aldehyde. After treatment with hydroxylamine hydrochloride and then sodium carbonate solution, aromatic aldehyde is converted to aldehyde oxime, which reacts with N-chlorosuccinimide (NCS) to afford aryl hydroximino chloride. 1,3-Dipole is eventually formed in situ while triethylamine (TEA) in DMF is added dropwise. Then 4-alkynyl sinomenine is added to provide the sinomenine isoxazole derivative via 1,3-dipolar cycloaddition reaction as the key step. All the target compounds are characterised by melting point, 1H NMR, 13C NMR, HRMS and FT-IR spectroscopy.

[Roles of G protein-coupled receptor 30 in the effects of genistein on apoptosis and cell cycle in human thyroid squamous cells SW579].

OBJECTIVE: Investigate the estrogen receptor expression in human thyroid squamous cell carcinoma SW579 and the effects of genistein on the apoptosis and cycle of SW579 and its mechnism. METHODS: The real-time PCR was applied to detect the expression of estrogen receptor(ER)α、ERß and G protein-coupled receptor(GPR)30 in human thyroid squamous cell line SW579; MTT was used to test the effect of genistein on cell proliferation in the SW579 cells before and after blocking GPR30; flow cytometry was explorited to measure the effect of genistein on the cell cycle and apoptosis in the SW579 was detected before and after blocking GPR30. RESULTS: The high concentration of genistein promoted the expression of ERß and GPR30 in the SW579 cells, but ERα was not expressed. The specific blocking of GPR30, the cell proliferation was aboviously inhibited by genistein in the SW579 cells and the cell apoptosis was markedly promoted after the GPR30 was blocked; The cell cycle was mainly blocked in G_2/M phase. CONCLUSION: Genistein can obiviously promote the cell proliferation in the SW579 cells, which may be related to the action of GPR30.

Identification and characterization of a pig FKBP5 gene with a novel expression pattern in lymphocytes and granulocytes.

The immunophilins are an important group of regulatory molecules in the immune system. FKBP5, expressed throughout mammals and in fish and birds, functions in both physiological and pathogenic pathways, including innate immunity and steroid-based diseases. In this study, we cloned the first porcine FKBP5 from Rongchang pig by the rapid amplification of cDNA ends technique. The full-length cDNA is 4097 bp, with an open reading frame of 1371 bp that codes for a 457-aa protein. Western blotting detected the porcine FKBP5 protein at highest levels in thymus, followed by spleen and lung. Immunohistochemistry detected the porcine FKBP5 protein in lymphocytes and granulocytes of the blood, and flow cytometry identified greater expression in unactivated (vs. activated) T lymphocytes. Finally, the expression level of porcine FKBP5 in the granulocytes was found to decline significantly from the time of birth to one-year-old. These collective data suggest that the newly identified porcine FKBP5 may function in activation of T cells in pig and in innate immunity in the newborn pig in particular.

[Effects of genistein on proliferation and invasion of human thyroid squamous cell carcinoma SW579 cells].

OBJECTIVE: To study the effect and mechanism of genistein on the proliferation, adhesion, invasion and migration of SW579 cells. METHODS: MTT assay was used to detect the effect of genistein on cell proliferation of the thyroid carcinoma cell line( SW579). Cell-matrix adhesion and transwell invasion assay were used to measure the impact of genistein on the cell adhesion, invasion and migration. Real-time PCR assay was used to detect the effect of genistein on the level of MMP2 mRNA. RESULTS: Compared with the control groups, genistein treated groups can significantly promote the proliferation of SW579 cells. However, genistein can obiviously inhibited the adhesion, invasion and migartion of SW579 cells. And genistein can dramatically decreased MMP2 mRNA expression in SW579 cells. CONCLUSION: In the study, genistein could effectively inhibit the invasion and migration ability of the SW579 cells in vitro, and the reduction of MMP-2 expression may play an important role in this process.

Impact of perinatal exposure to acetaminophen on hepatocellular metabolic function in offspring.

Acetaminophen (APAP), an over the counter (OTC) medication, is widely used in antipyretic treatment. Although the risk of dose-dependent cytotoxicity has been known, the potential effect of perinatal exposure to acetaminophen on metabolic function in offspring remains uninvestigated. Therefore, we established a prenatally APAP-exposed pregnancy mouse model to assess the possible adverse effect on liver metabolic function in offspring. Biochemical assays were applied in analysis of basic metabolic parameters in postnatal mice. Further, immunoblotting assay was used to assess the expressions of insulin receptor ß (IRß), insulin receptor substrate 1 (IRS1), phospho-Akt and phospho-GSK-3ß proteins in liver cells. In addition, hepatic glucose transporter 2 (GLUT2) immunoactivity was determined by using immunohistochemistry staining. Compared with untreated postnatal mice, APAP-exposed offspring induced impaired glucose metabolism, increased plasma insulin level, and reduced liver glycogen content. In addition, APAP exposure decreased the expressions of IRS1 and phospho-GSK-3ß, phospho-AKT proteins and down-regulated the level of glucose-import regulator GLUT2 in the liver. Taken together, our preliminary findings indicate that perinatal APAP exposure-impaired hepatic glucose metabolism in offspring may be associated with disturbance of insulin-dependent AKT signaling in the liver.

[The change of cancer-related genes expression profile in Nthy-ori-3-1 cell induced by the pesticide amitrole].

OBJECTIVE: To explore the possible mechanism of amitrole causing thyroid tumor in Nthy-ori-3-1 cell by differential expression microarray analysis. METHODS: After the Nthy-ori-3-1cells were treated with 1 ~ 100 g / m L amitrole for 24 h, and the effect of amitrole on the proliferation of the cells was detected by MTT assay. Then cells were treated with 100 g / m L amitrole for 24 h, and the differential expression microarray was tested. The microarray results was analyzed by GO analysis and pathway analysis. The microarray results were verified by real-time quantitative PCR. RESULTS: MTT results showed that amitole had no significant effect on the proliferation of Nthy-ori-3-1 cells. Microarray results showed that 90( 55 up-regulated, 35 down regulated) genes were significantly changed. GO analysis showed that 43( 37 up-regulated, 6 down-regulated) of the 90 changed genes were related to biological processes, and 42( 37 up-regulated, 5down-regulated) were related to molecular function, and 44( 38 up-regulated, 6 downregulated) were related to cell components. Pathway results showed that 44 signalingpathways were influenced by the differentially expressed genes, and 10 of them were closely related to tumor. The qRT-PCR results were consistent with microarray results. wnt5 b, arnt2 and bmp2 genes were significantly related with multiple tumor-associated pathways. CONCLUSION: Amitrole may cause thyroid tumor by multiple signaling pathways, and bmp2, arnt2 and wnt5 b may beits major target genes.

Stimulating effect of a newly synthesized sulfonamido-based gallate on articular chondrocytes in vitro.

BACKGROUND: The phenotype of chondrocyte is easy to be lost when expanded in vitro by a process defined "dedifferentiation". Traditional growth factors such as transforming growth factor (TGF-ß1) are effective in preventing of dedifferentiation, but high costs and loss of activity limited their use. It is of significance to find substitutes which can reduce dedifferentiation and preserve chondrocytes phenotype to ensure sufficient differentiated cells for further study. METHODS: We synthesized new type of sulfonamido-based gallates named ZXHA-C and investigated its effect on primary articular chondrocytes of rats. After preliminary screening by cytotoxicity test, ZXHA-C of 1.06 × 10-8, 1.06 × 10-7 and 1.06 × 10-6M were chosen for further studies. Cell proliferation, morphology, viability, GAG synthesis and cartilage specific gene expression were detected. Also the effects of ZXHA-C on Wnt/ß-catenin signaling pathway were investigated. RESULTS: ZXHA-C could significantly promote chondrocytes growth. And it could enhance ECM synthesis by up-regulating expression levels of cartilage specific markers like aggrecan, collagen II and Sox9. Expression of collagen I which marked chondrocytes dedifferentiation was also significantly down-regulated after treated by ZXHA-C. Further exploration of the molecular mechanism indicated that ZXHA-C activated the Wnt/ß-catenin signal pathway in chondrocytes, as evidenced by up-regulated gene expression of ß-catenin, Wnt-4, cyclin D1 and Frizzled-2 and decreased glycogen synthase kinase 3ß (GSK-3ß). Among the various concentrations, ZXHA-C of 1.06 × 10-7 M showed the best performance, which was close to positive control (group with TGF-ß1). CONCLUSION: ZXHA-C might be potential a novel agent for the maintenances of chondrocytes phenotype.

[Whole-genome sequencing and analysis of inosine- producing strain Bacillus subtilis ATCC 13952].

OBJECTIVE: Bacillus subtilis ATCC 13952 is an inosine-producing strain. In order to study the mechanisms of inosine accumulation and offer help for molecular breeding, it is necessary to uncover the genome sequence of ATCC 13952. METHODS: Whole-genome sequencing of ATCC 13952 is carried out by Solexa and Sanger sequencing. Genome assembly, gene prediction and functional annotation, GO/COG cluster analysis and synteny analysis are done using relevant software. RESULTS: The complete genomic information of Bacillus subtilis ATCC 13952 is contained on a single circular chromosome of 3876276 bp with an average GC content of 45.8%. The genome sequence is deposited in the GenBank under the accession number CP009748. Comparative genomic analysis shows that ATCC 13952 should have significant genomic synteny with other Bacillus subtilis strains. On the other hand, some point mutation and deletions occurred in purine metabolism-related genes between ATCC 13952 and the standard strain. CONCLUSION: The results of this study will provide a theoretical basis for subsequent further molecular breeding.

[Mechanism of the effects of amitrole on the thyroglobulin in Fischer rat thyroid follicle-5 cell].

OBJECTIVE: The mechanisms of the effects of amitrole on the thyroglobulin (TG) was investigated in Fischer rat thyroid follicle-5 cell (FRTL-5 cells) and in the medium. METHODS: FRTL-5 cells were treated with 1, 10 and 100 microg/ml amitrole, and cytotoxicity was tested by 3H-TdR. The effects of amitrole on TG and thyroid transcription factor 1(TTF-1) in FRTL-5 cells were analyzed by RIA and ICC. And the TSHR in FRTL-5 cells was examined by Immunofluorescence analysis. RESULTS: 1,10 and 100 microg/ml amitrole had no significant cytotoxicity to the FRTL-5 cells (P > 0.05). The concentration of TG in the culture was decreased by 10 and 100 microg/ml amitrole (P < 0.05), and the concentration of TG in the cells was decreased by 100 microg/ml (P < 0.05) but the TTF-1 in the cells were not obviously changed (P > 0.05). The TSHR in the surface of FRTL-5 cells was significantly decreased by all doses of amitrole (P < 0.01). CONCLUSION: The influence of amitrole on TG of FRTL-5 cells may be related to significantly reduce TSHR in the surface of FRTL-5 cells.

[Effect of different fertilizing levels on yields and efficiency components of Pinellia ternata from Sichuan].

OBJECTIVE: The effect of the different fertilizing levels on yields and main active components of Pinellia ternata from Sichuan was studied under the cultivated condition, in order to provide theoretical basis for the standardized cultivation. METHOD: Using one of the wild populations of P. ternate from Sichuan as tested material, the experiment was performed with orthogonal designing methods L25 (5(3)). During growth and development period, agronomic traits such as number of sprouting, inflorescence and bulblets were counted. After harvesting, main chemical compositions, growth and proliferation rates were determined. RESULT: In different fertilizing levels, the P. ternata from Sichuan showed the same growth rhythm, though there were significant difference (P < 0.05) among the average emergence rate,whereas extremely significant difference (P < 0.01) were detected among the average ratio of bolting, the average bulbils, individual growth rate, individual proliferation rate, beta-sitosterol and alkaloid content under different fertilizing levels. Nitrogenous fertilizer that affected the content of alkaloids and beta-sitosterol were extremely significant (P < 0.01), whereas phosphate and potassium fertilizer had no significant effect. The effect of fertilizer factor and inter effects on beta-sitosterol showed no significant effect, but have influence on other indexes. CONCLUSION: The optimum fertilizer composition was 315 kg x hm(-2) of nitrogen, 225 kg x hm(-2) of P2O5 and 270 kg x hm(-2) of K2O.

The effects of amitrole on thyroglobulin and iodide uptake in FRTL-5 cells.

Thyroid is a frequent target for endocrine effects of pesticides. Thyroglobulin (TG) and iodide uptake are crucial to thyroid hormone synthesis and may be targets of thyroid-disrupting chemicals. In our study, thyroid follicular FRTL-5 cells were treated with amitrole, an inhibitor of the thyroid peroxidase (TPO), and the effects on TG and total iodide uptake were observed. The results showed that 1-100 mg/L amitrole had no marked effects on FRTL-5 cell proliferation and DNA synthesis. However, it significantly increased the transcription of tg gene and inhibited the total iodide uptake. And 10-100 mg/L amitrole significantly decreased TG in the culture medium. The data suggests amitrole may disrupt the expression and secretion of TG and iodide uptake.