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1.
Beijing Da Xue Xue Bao Yi Xue Ban ; 54(4): 658-662, 2022 Aug 18.
Article in Chinese | MEDLINE | ID: mdl-35950388

ABSTRACT

OBJECTIVE: To investigate the change trends of sperm quality with seasonal variations among the volunteers of sperm donors in Beijing area, as well as the relationship between two parameters. METHODS: Semen data from the volunteers of sperm donors in Human Sperm Bank of Peking University Third Hospital were collected using a retrospective study method. The subjects were divided into 4 seasonal groups based on the lunar solar terms and the time of sperm donation. The data were assessed to find whether there were differences in semen parameters among different seasonal groups, and to analyze the change trends and the influence of seasonal factors on semen parameters. RESULTS: A total of 21 174 semen parameter data were analyzed. Firstly, to analyze all data as a whole, in spring, summer, autumn and winter groups, sperm concentration was (106.04±59.67)×106/mL, (97.61±47.41)×106/mL, (100.18±47.17)×106/mL, (100.59±38.68)×106/mL, respectively, and the spring group was significantly higher than the other 3 seasonal groups (P < 0.001); proportion of progressive motility sperm (PR) was 56.49%±12.76%, 58.02%±13.65%, 58.05%±12.36%, and 57.66%±12.61%, respectively, spring group was lower than the other three seasonal groups, and summer group was better among the latter (P < 0.001). There was no difference in normal rate of sperm morphology among the four seasonal groups. The qualified rate of sperm donors in the winter group was higher than that in the other three seasons groups (P < 0.01), while the qualified rate in the summer group was lower than that in the other three seasons groups. In addition, the semen parameters of the volunteers during the screening period and the official sperm donation period were analyzed respectively, which revealed that sperm concentration of spring group was higher than that of summer and winter groups, and PR was lower than that of summer and autumn groups. On account of the semen parameters of official sperm donation period, multiple linear regression analysis found that season was the main factor affecting sperm concentration, the average sperm concentration in spring group was about 6×106/mL higher than in winter group, but PR was 2.9% lower in spring group compared with autumn group (all P < 0.05). CONCLUSION: Season was the influencing factor of semen quality of sperm donors in Beijing area. We recommend spring and winter may be the preferred seasons for sperm donation.


Subject(s)
Semen Analysis , Semen , Humans , Male , Retrospective Studies , Seasons , Sperm Count , Sperm Motility , Spermatozoa
2.
Zhonghua Xin Xue Guan Bing Za Zhi ; 48(4): 323-328, 2020 Apr 24.
Article in Chinese | MEDLINE | ID: mdl-32370484

ABSTRACT

Objective: To investigate whether inflammatory factor tumor necrosis factor-α (TNF-α) is involved in the electrical remodeling of cardiomyocytes by regulating ultra-rapid delayed rectifier K(+) current (I(kur)) and the role of Src kinase. Methods: H9c2 cells, embryonic cardiomyocytes of rat, were cultured in Dulbecco's modified Eagle's medium (DMEM) and atrium-derived HL-1 cells were cultured in Claycomb medium. Both H9c2 and HL-1 cells were cultured at 37 ℃ with 5% CO(2). Cells cultured in normal conditions without additional treatment served as control group. Experimental groups were treated with different concentration of TNF-α (25 or 50 or 100 ng/ml) for 24 hours. To study whether Src specific inhibitor PP1 could abrogate the effect of TNF-α, cells were pre-treated with 10 µmol/L PP1 for 1 hour, followed by TNF-α (100 ng/ml) for 24 hours. Western blot and the whole cell patch clamp technique were used to detect the protein expression of Kv1.5 and Src and I(kur) in each group. Results: (1) In H9c2 cells, high concentration of TNF-α treatment (100 ng/ml) significantly reduced the Kv1.5 protein expression compared with control group and TNF-α 25 ng/ml group (both P<0.05). Compared with control group, the expression of p-Src protein was higher in 25 ng/ml, 50 ng/ml, 100 ng/ml TNF-α group (all P<0.05), but there was no statistical difference in the expression of Src protein among groups (P>0.05). In addition, the current density of I(kur) was decreased in 50 ng/ml, 100 ng/ml TNF-α group (both P<0.05). Furthermore, the expression of Kv1.5 protein and the current density of I(kur) were increased in PP1+TNF-α group compared with TNF-α 100 ng/ml group (both P<0.05). There was no statistical difference in the expression of Kv1.5 protein and the current density of I(kur) between the control group and PP1+TNF-α group (both P>0.05). (2) In atrium-derived HL-1 cells, the expression of Kv1.5 protein was reduced in 100 ng/ml TNF-α group compared with control group and TNF-α 25 ng/ml group (both P<0.01). In addition, the expression of p-Src protein was increased in TNF-α 100 ng/ml group compared with control group (P<0.05), but there was no statistical difference in the protein expression of Src among groups (P>0.05). The expression of Kv1.5 protein was increased in PP1+TNF-α group compared with TNF-α 100 ng/ml group (P<0.05). Conclusion: TNF-α is involved in the pathogenesis of atrial fibrillation, probably via decreasing I(kur) current density in atrium-derived myocytes through the activation of Src kinase.


Subject(s)
Down-Regulation , Myocytes, Cardiac , Animals , Heart Atria , Rats , Tumor Necrosis Factor-alpha , src-Family Kinases
3.
Scand J Immunol ; 84(2): 86-94, 2016 Aug.
Article in English | MEDLINE | ID: mdl-27146476

ABSTRACT

Anti-CD45RB monoclonal antibody (anti-CD45RBmAb), as a new immune tolerance inducer, may inhibit T cell proliferation and induce immune tolerance through competitive combination with CD45RB on the T cell surface, which blocks the conduction of activation signals. However, how anti-CD45RBmAb plays its role on T lymphocyte subsets during immunosuppression remains unclear. In this work, we investigate the effects of anti-CD45RBmAb on CD3(+) T lymphocyte both in vitro and in allogeneic heart transplant model in vivo. Interestingly, anti-CD45RBmAb could inhibit the proliferation of T cells, promote the transformation of T lymphocyte to Treg and Th2 cells, suppress the transformation to Th17 and Th1 cells, increase the number of Ts cells, decrease the number of Tm cells and thus play a role in immune inhibition and induction of immune tolerance.


Subject(s)
Antibodies, Monoclonal/therapeutic use , Graft Rejection/prevention & control , Heart Transplantation , T-Lymphocytes, Regulatory/drug effects , Th2 Cells/drug effects , Animals , Cell Differentiation/drug effects , Cell Proliferation/drug effects , Cells, Cultured , Humans , Immune Tolerance/drug effects , Leukocyte Common Antigens/immunology , Lymphocyte Activation/drug effects , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Models, Animal , T-Lymphocytes, Regulatory/immunology , Th2 Cells/immunology
4.
Springerplus ; 5: 372, 2016.
Article in English | MEDLINE | ID: mdl-27066380

ABSTRACT

An ideal magnetic rail should provide a homogeneous magnetic field along the longitudinal direction to guarantee the reliable friction-free operation of high temperature superconducting (HTS) maglev vehicles. But in reality, magnetic field inhomogeneity may occur due to lots of reasons; the joint gap is the most direct one. Joint gaps inevitably exist between adjacent segments and influence the longitudinal magnetic field homogeneity above the rail since any magnetic rails are consisting of many permanent magnet segments. To improve the running performance of maglev systems, two new rail joints are proposed based on the normal rail joint, which are named as mitered rail joint and overlapped rail joint. It is found that the overlapped rail joint has a better effect to provide a competitive homogeneous magnetic field. And the further structure optimization has been done to ensure maglev vehicle operation as stable as possible when passing through those joint gaps. The results show that the overlapped rail joint with optimal parameters can significantly reduce the magnetic field inhomogeneity comparing with the other two rail joints. In addition, an appropriate gap was suggested when balancing the thermal expansion of magnets and homogenous magnetic field, which is considered valuable references for the future design of the magnetic rails.

5.
Genet Mol Res ; 15(1)2016 Feb 22.
Article in English | MEDLINE | ID: mdl-26910002

ABSTRACT

5-Aminolevulinate synthase 1 (ALAS1) is the first enzyme in the heme biosynthetic pathway and is upregulated in follicular tissue during the early stages of ovulation. In this study, we isolated the complete coding sequence of the porcine ALAS1 gene and its 2-9 intron sequence, identified a single nucleotide polymorphism (SNP; T/C) in intron 9, and developed a PCR-MspI-restriction fragment length polymorphism genotyping assay. Association of the SNP with litter size was assessed in two populations [purebred Large White and the experimental synthetic (DIV) line]. Statistical analysis demonstrated that for total number of piglets born (TNB) in all parities, pigs with the CC genotype had an additional 0.68 and 0.74 piglets compared to the TC and TT animals (P < 0.05) in the DIV line, respectively. Purebred Large White sows inheriting the CC and TC genotypes gave birth to an additional 0.96 and 0.70 piglets compared to the TT animals (P < 0.05) in all parities, respectively. In addition, for TNB in all parities, a significant additive effect of 0.48 ± 0.23 and 0.37 ± 0.17 piglets/ litter was detected in sows of both populations (P < 0.05), respectively. The highest levels of ALAS1 gene expression were observed in isolated ovarian granulosa cells 2 and 12 h after stimulation with pregnant mare serum gonadotropin human chorionic gonadotropin, which represents the time of follicular development and ovulation, respectively. Therefore, the ALAS1 gene was significantly associated with litter size in two populations and could be a useful molecular marker for the selection of increasing litter size in pigs.


Subject(s)
5-Aminolevulinate Synthetase/genetics , Litter Size/genetics , Parity/genetics , Polymorphism, Single Nucleotide , 5-Aminolevulinate Synthetase/metabolism , Animals , Female , Introns , Ovary/metabolism , Ovulation/genetics , Pregnancy , Swine
6.
J Nanosci Nanotechnol ; 15(3): 2384-8, 2015 Mar.
Article in English | MEDLINE | ID: mdl-26413672

ABSTRACT

Through the pyrolysis of acetylene at 250 °C, large quantities of carbon nanofiber bundles (CNFBs), curved carbon nanofibers (CCNFs) and helical carbon nanofibers (HCNFs) can be synthesized selectively by controlling the Fe:Cu molar ratio of Fe-Cu nanoparticles. In this study, the systematic experimental results indicated that the Cu content in the Fe-Cu nanoparticles and pyrolysis temperature had great impact on the yield and structure of the final samples. Moreover, the transmission electron microscopic observation indicated that the catalyst nanoparticles were enwrapped tightly by graphite layers, and the obtained HCNFs show good magnetic property. Compared to the methods reported in the literature, the approach described herein has the advantages of being simple, low-cost, and environment-friendly. It is suitable for the controllable and mass production of CNFBs, CCNFs and HCNFs.

7.
Genet Mol Res ; 13(1): 1195-202, 2014 Feb 27.
Article in English | MEDLINE | ID: mdl-24634176

ABSTRACT

The titin immunoglobulin domain (TTID) protein localizes to the Z line in muscle and binds to alpha-actinin and gamma-filamin. It plays an indispensable role in stabilizing and anchoring of thin filaments. In this study, the 5'-regulatory region of the porcine TTID gene was analyzed with bioinformatic methods. Another objective of this study was to further investigate the polymorphism in the intron 6 of the porcine TTID gene. We determined allele frequency among six Chinese porcine purebreds. The polymorphisms were genotyped in a population of 280 F2 pigs representing two Large White x Meishan reference families. Different TTID genotypes were significantly associated with carcass traits, including skin percentage (P < 0.05), loin eye area (P < 0.05), and average skin thickness (P < 0.01). Our study will continue to lay the groundwork for further investigations into the detailed function of the porcine TTID gene.


Subject(s)
Body Composition , Connectin/genetics , Genetic Association Studies/methods , 5' Flanking Region , Animal Husbandry , Animals , Base Sequence , Gene Frequency , Genotype , Least-Squares Analysis , Molecular Sequence Data , Polymorphism, Single Nucleotide , Quantitative Trait Loci , Swine
8.
Int J Tuberc Lung Dis ; 18(4): 492-8, 2014 Apr.
Article in English | MEDLINE | ID: mdl-24670708

ABSTRACT

OBJECTIVE: To investigate whether tobacco smoking increases the risk of tuberculosis (TB) recurrence and identify factors associated with TB recurrence among adults who had successfully completed anti-tuberculosis treatment in Taipei, Taiwan. METHODS: Recurrence was defined as a new clinical or microbiological diagnosis of TB requiring the start of a new course of treatment in a patient who had satisfactorily completed treatment for a previous TB episode. Cox proportional hazard models were used to calculate adjusted hazard ratios (aHRs) for recurrence. RESULTS: We followed 5567 adults for recurrence after successful anti-tuberculosis treatment. The mean age was 58.5 years; 62.9% were male. Overall, 84 (1.5%) had a recurrence of TB during follow-up. The incidence of TB recurrence was 4.9 episodes/1000 person-years of follow-up. Cox proportional hazards regression showed that after controlling for other variables, the risk of TB recurrence among subjects who smoked >10 cigarettes a day was double that of never/former smokers. Other independent risk factors significantly associated with TB recurrence were homelessness (aHR 3.75, 95%CI 1.17-12.07), presence of comorbidities (aHR 2.66, 95%CI 1.22-5.79) and a positive acid-fast bacilli smear (aHR 2.27, 95%CI 1.47-3.49). CONCLUSION: Smoking >10 cigarettes a day was significantly associated with TB recurrence. To reduce the risk of recurrence, we recommend including effective measures of smoking cessation in TB control programmes, as recommended by the World Health Organization Stop TB Strategy.


Subject(s)
Antitubercular Agents/therapeutic use , Smoking/adverse effects , Tuberculosis, Pulmonary/drug therapy , Adolescent , Adult , Aged , Aged, 80 and over , Female , Humans , Kaplan-Meier Estimate , Male , Middle Aged , Multivariate Analysis , Proportional Hazards Models , Recurrence , Retrospective Studies , Risk Assessment , Risk Factors , Smoking/epidemiology , Taiwan/epidemiology , Time Factors , Treatment Outcome , Tuberculosis, Pulmonary/diagnosis , Tuberculosis, Pulmonary/epidemiology , Young Adult
9.
Braz J Med Biol Res ; 46(9): 746-51, 2013 Sep.
Article in English | MEDLINE | ID: mdl-24036910

ABSTRACT

Macrophage migration inhibitory factor (MIF), a pleiotropic cytokine, plays an important role in the pathogenesis of atrial fibrillation; however, the upstream regulation of MIF in atrial myocytes remains unclear. In the present study, we investigated whether and how MIF is regulated in response to the renin-angiotensin system and oxidative stress in atrium myocytes (HL-1 cells). MIF protein and mRNA levels in HL-1 cells were assayed using immunofluorescence, real-time PCR, and Western blot. The result indicated that MIF was expressed in the cytoplasm of HL-1 cells. Hydrogen peroxide (H2O2), but not angiotensin II, stimulated MIF expression in HL-1 cells. H2O2-induced MIF protein and gene levels increased in a dose-dependent manner and were completely abolished in the presence of catalase. H2O2-induced MIF production was completely inhibited by tyrosine kinase inhibitors genistein and PP1, as well as by protein kinase C (PKC) inhibitor GF109203X, suggesting that redox-sensitive MIF production is mediated through tyrosine kinase and PKC-dependent mechanisms in HL-1 cells. These results suggest that MIF is upregulated by HL-1 cells in response to redox stress, probably by the activation of Src and PKC.


Subject(s)
Hydrogen Peroxide/pharmacology , Intramolecular Oxidoreductases/drug effects , Macrophage Migration-Inhibitory Factors/drug effects , Myocytes, Cardiac/metabolism , Oxidants/pharmacology , Protein Kinase C/metabolism , src-Family Kinases/metabolism , Angiotensin II/metabolism , Animals , Blotting, Western , Cell Line , Immunohistochemistry , Intramolecular Oxidoreductases/genetics , Macrophage Migration-Inhibitory Factors/genetics , Mice , Microscopy, Confocal , Oxidative Stress/physiology , Protein Kinase Inhibitors/pharmacology , Real-Time Polymerase Chain Reaction , Renin-Angiotensin System/physiology
10.
Int J Tuberc Lung Dis ; 17(10): 1310-6, 2013 Oct.
Article in English | MEDLINE | ID: mdl-24025383

ABSTRACT

OBJECTIVE: To identify factors associated with death before the start of anti-tuberculosis treatment, and early and late during treatment, among adult Taiwanese with culture-positive pulmonary tuberculosis (PTB). METHOD: All adult culture-positive PTB patients in Taipei, Taiwan, were included in a retrospective cohort study in 2005-2010. RESULTS: Of 4438 patients (mean age 64.6 years, 70.6% male), 76.8% were successfully treated, 5.4% died before start of treatment, 9.0% died within 8 weeks of treatment initiation and 8.8% died >8 weeks after treatment initiation. After controlling for potential confounders, age ≥ 65 years and male sex were associated with higher risks of death at all time periods investigated. High school education or higher reduced the risk of death before the start of and during treatment, while unemployment increased the risk of mortality during treatment. Cavity on chest X-ray and positivity for acid-fast bacilli were associated with lower risk of mortality before the start of treatment. CONCLUSION: To lower mortality among adult culture-positive PTB patients, it is imperative for clinicians to maintain high awareness of TB and provide more intensive care early, especially for men, the elderly and people with lower socio-economic status (e.g., the unemployed and less educated).


Subject(s)
Antitubercular Agents/therapeutic use , Tuberculosis, Pulmonary/drug therapy , Adolescent , Adult , Age Factors , Aged , Aged, 80 and over , Antitubercular Agents/administration & dosage , Cohort Studies , Educational Status , Female , Follow-Up Studies , Humans , Male , Middle Aged , Prognosis , Retrospective Studies , Risk Factors , Sex Factors , Socioeconomic Factors , Taiwan , Time Factors , Treatment Outcome , Tuberculosis, Pulmonary/mortality , Young Adult
11.
Braz. j. med. biol. res ; 46(9): 746-751, 19/set. 2013. graf
Article in English | LILACS | ID: lil-686569

ABSTRACT

Macrophage migration inhibitory factor (MIF), a pleiotropic cytokine, plays an important role in the pathogenesis of atrial fibrillation; however, the upstream regulation of MIF in atrial myocytes remains unclear. In the present study, we investigated whether and how MIF is regulated in response to the renin-angiotensin system and oxidative stress in atrium myocytes (HL-1 cells). MIF protein and mRNA levels in HL-1 cells were assayed using immunofluorescence, real-time PCR, and Western blot. The result indicated that MIF was expressed in the cytoplasm of HL-1 cells. Hydrogen peroxide (H2O2), but not angiotensin II, stimulated MIF expression in HL-1 cells. H2O2-induced MIF protein and gene levels increased in a dose-dependent manner and were completely abolished in the presence of catalase. H2O2-induced MIF production was completely inhibited by tyrosine kinase inhibitors genistein and PP1, as well as by protein kinase C (PKC) inhibitor GF109203X, suggesting that redox-sensitive MIF production is mediated through tyrosine kinase and PKC-dependent mechanisms in HL-1 cells. These results suggest that MIF is upregulated by HL-1 cells in response to redox stress, probably by the activation of Src and PKC.


Subject(s)
Animals , Mice , Hydrogen Peroxide/pharmacology , Intramolecular Oxidoreductases/drug effects , Macrophage Migration-Inhibitory Factors/drug effects , Myocytes, Cardiac/metabolism , Oxidants/pharmacology , Protein Kinase C/metabolism , src-Family Kinases/metabolism , Angiotensin II/metabolism , Blotting, Western , Cell Line , Immunohistochemistry , Intramolecular Oxidoreductases/genetics , Microscopy, Confocal , Macrophage Migration-Inhibitory Factors/genetics , Oxidative Stress/physiology , Protein Kinase Inhibitors/pharmacology , Real-Time Polymerase Chain Reaction , Renin-Angiotensin System/physiology
12.
Transplant Proc ; 43(7): 2721-7, 2011 Sep.
Article in English | MEDLINE | ID: mdl-21911152

ABSTRACT

Currently lifelong immunosuppression is required for organ transplant recipients. Anti-CD45RB monoclonal antibody (mAb) prolongs graft survival by mechanisms that are not yet clear. Therefore, we investigated the role of T and dendritic cells (DC) in islet allografts treated with anti-CD45RB mAb after transplantation of 200 allogeneic islets (BALB/c mouse) under the kidney capsules of diabetic C57BL/6 mice treated with intraperitoneal injections of 100 µg of anti-CD45RB mAb on days 0, 1, 3, 5, and 7. We observed a tilt of the ratios of Th1/Th2 and Tc1/Tc2 to Th2 and Tc2. The numbers of naïve and memory T cells were down-regulated in peripheral blood after transplantation. In addition, the maturation, endocytosis, and interleukin-12 secreted by DC derived from bone marrow cells was suppressed in recipient mice. Therefore, anti-CD45RB mAb alleviated, rejection by suppressive effects on T-lymphocyte subsets and DC.


Subject(s)
Antibodies, Monoclonal/immunology , Dendritic Cells/immunology , Islets of Langerhans/immunology , Leukocyte Common Antigens/immunology , T-Lymphocytes/immunology , Animals , Diabetes Mellitus, Experimental/immunology , Flow Cytometry , Immunologic Memory , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Streptozocin , Transplantation, Homologous
13.
Genetika ; 47(4): 537-42, 2011 Apr.
Article in English | MEDLINE | ID: mdl-21675243

ABSTRACT

Imprinted genes play significant roles in the regulation of fetal growth and development, function of the placenta, and maternal nurturing behaviour in mammals. At present, few imprinted genes have been reported in pigs compared to human and mouse. In order to increase understanding of imprinted genes in swine, a polymorphism-based approach was used to assess the imprinting status of three porcine genes in 12 tissue types, obtained from F1 pigs of reciprocal crosses between Rongchang and Landrace pure breeds. In contrast to human and mouse homologues, porcine PPP1R9A was not imprinted, and was found to be expressed in all tissues examined. The expression of porcine NAP1L5 was detected in pituitary, liver, spleen, lung, kidney, stomach, small intestine, skeletal muscle, fat, ovary, and uterus, but undetectable in heart. Furthermore, porcine NAP1L5 was paternally expressed in the tissues where it's expression was observed. For PEG3, pigs expressed the paternal allele in skeletal muscle, liver, spleen, kidney, and uterus, but biallele in heart, lung, fat, stomach, small intestine, and ovary. Our data indicate that tissue distribution of the three gene differs among mammals, and the imprinting of NAP1L5 and PEG3 is well conserved.


Subject(s)
Genomic Imprinting/genetics , Sus scrofa/genetics , Alleles , Animals , Cattle , Gene Expression Regulation, Developmental/genetics , Humans , Kruppel-Like Transcription Factors/genetics , Kruppel-Like Transcription Factors/metabolism , Mice , Microfilament Proteins/genetics , Microfilament Proteins/metabolism , Nerve Tissue Proteins/genetics , Nerve Tissue Proteins/metabolism , Organ Specificity , Polymorphism, Genetic
14.
Mol Biol Rep ; 38(2): 827-32, 2011 Feb.
Article in English | MEDLINE | ID: mdl-20376701

ABSTRACT

In this study, the expression profiling of three troponin I isoforms (TNNI1, TNNI2 and TNNI3) was investigated in two pig breeds differing in muscularity (Yorkshire and Meishan) at six stages (fetal 60 days and postnatal 3, 35, 60, 120, and 180 days) and three types of muscles (longissimus dorsi muscle, LD; semitendinosus, ST; cardiac muscle, CM) using relative real-time quantitative PCR. Significant differences of troponin I expression in three muscles were found between Yorkshire and Meishan breeds at some stages. The expression peak of TNNI1 and TNNI2 in LD and ST was at postnatal 35 or 60 days in Yorkshire and at postnatal 120 or 180 days in Meishan pigs, while it occurred in CM at postnatal 3 days in two pig breeds. The relative expression values of TNNI1 and TNNI2 were significantly higher in LD than ST at most of stages after birth. The expression ratio of TNNI2 versus TNNI1 favoured TNNI2 expression in ST and LD, but on the contrary in CM. The expression peak of TNNI3 occurred at postnatal 60 and 120 days in Yorkshire and Meishan pigs, respectively. TNNI1 and TNNI3 were co-expressed in CM during the fetal and earlier stages after birth.


Subject(s)
Gene Expression Profiling , Heart/embryology , Muscles/metabolism , Myocardium/cytology , Troponin I/genetics , Animals , DNA Primers/genetics , RNA/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Species Specificity , Swine , Time Factors , Troponin I/biosynthesis
15.
Mol Biol Rep ; 38(1): 103-14, 2011 Jan.
Article in English | MEDLINE | ID: mdl-20333549

ABSTRACT

Nuclear receptor subfamily 4, group A, member 1 (NR4A1), other aliase NGFI-B, is an immediate-early gene that encodes an orphan nuclear receptor, which play a potential role in the ovulatory process. In this study, a 4,870 bp fragment covered the complete coding region (CDS) and its unique intron sequences of porcine NR4A1 gene was obtained. The reverse transcriptase-polymerase chain reaction (RT-PCR) indicated that NR4A1 was highly expressed in ovary, uterus, kidney, heart but at very low level in oviduct and not expressed in other tissues. Compared the sequence of CDS and its unique intron of Large White and Meishan pigs, a A/G mutation in intron 5 was found and a PCR-Dde1-RFLP genotyping assay was developed. Association of the SNP and litter size was assessed in two populations [purebred Large White and an experimental synthetic Line (DIV) sows]. Statistical analysis demonstrated that, in the first parity, AG animals in experimental synthetic Line (DIV) sows had 1.805 more piglets born compared to the GG animals (P<0.05). For all parities, in the purebred Large White pigs, those with the GG genotype had an additional 0.877 piglets born and 0.780 piglets born alive compared to the AA animals (P<0.05), those with the AG genotype had additional 0.780 piglets born compared to the AA animals (P<0.05). In addition, significant additive effect of 0.438±0.182 piglets/litter and 0.368±0.165 piglets/litter on piglets born and piglets born alive were detected in the purebred Large White lines (P<0.05), respectively. Therefore, NR4A1 gene was significantly associated with litter size in two populations and could be a useful molecular marker in selection for increasing litter size in pigs.


Subject(s)
Gene Expression Regulation , Genetic Association Studies , Nuclear Receptor Subfamily 4, Group A, Member 1/genetics , Quantitative Trait, Heritable , Reproduction/genetics , Sus scrofa/genetics , Amino Acid Sequence , Animals , Base Sequence , Breeding , Cloning, Molecular , Conserved Sequence/genetics , Gene Expression Profiling , Gene Frequency/genetics , Genetic Variation , Humans , Mice , Molecular Sequence Data , Nuclear Receptor Subfamily 4, Group A, Member 1/chemistry , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Protein Structure, Tertiary , Sequence Analysis, DNA
16.
Mol Biol Rep ; 38(1): 267-74, 2011 Jan.
Article in English | MEDLINE | ID: mdl-20336382

ABSTRACT

Skeletal muscle and kidney enriched inositol phosphatase (SKIP) was identified as a 5'-inositol phosphatase that hydrolyzes PI(3,4,5)P3 to PI(3,4)P2 that negatively regulates insulin-induced phosphatidylinositol 3-kinase signaling in skeletal muscle. In this study, we obtained a 1575-bp mRNA sequence of porcine SKIP that included the full coding region encoding a protein of 450 amino acids. With the use of comparative mapping, we mapped this gene to SSC12 q1.3, where many QTLs affect Backfat thickness at 10th rib, carcass yield, the number of muscle fibers, and ham weight traits. As a candidate gene for growth and carcass traits, a novel single nucleotide polymorphism in exon 12 (G>A) was detected by PCR-RFLP. The results showed that the GG genotype had higher skin percentage (SP), carcass length to first spondyle (CL1), carcass length to first rib (CL2), but lower intramuscular fat (IMF) as compared with genotype AG (P<0.05), and allele G seemed to be associated with an increase in the growth trait. Porcine SKIP was expressed abundantly in skeletal muscle tissue and was transcriptionally upregulated during skeletal muscle differentiation. Analysis of the porcine SKIP promoter sequence demonstrated that MyoD was involved in regulating SKIP mRNA expression in myotubes, partly via the cis-acting elements in SKIP promoter. In summary, we suggested that SKIP might play a role in the regulation of skeletal muscle development in pigs.


Subject(s)
Gene Expression Regulation, Developmental , Muscle Development/genetics , Muscle, Skeletal/enzymology , Muscle, Skeletal/growth & development , MyoD Protein/metabolism , Phosphoric Monoester Hydrolases/genetics , Sus scrofa/growth & development , Animals , Base Sequence , Breeding , Cell Differentiation/genetics , Cell Line , Cloning, Molecular , Exons/genetics , Gene Frequency/genetics , Genetic Association Studies , Genetic Loci/genetics , Genotype , Humans , Luciferases/metabolism , Mice , Myoblasts/cytology , Myoblasts/metabolism , Phosphoric Monoester Hydrolases/metabolism , Polymorphism, Genetic , Promoter Regions, Genetic/genetics , Sus scrofa/genetics , Transcription, Genetic , Up-Regulation/genetics
17.
Mol Biol Rep ; 38(6): 3725-9, 2011 Aug.
Article in English | MEDLINE | ID: mdl-21104015

ABSTRACT

MicroRNAs (miRNAs) are 20-25 nt, endogenous non-coding RNA molecules that act by binding to the complementary sequence of target messenger RNAs. Many evidences showed that miRNAs were involved in the process of germ proliferation and differentiation. In the present study, miR-27a gene was selected as a candidate gene for litter size due to its biological function, its location near a mummified pigs QTL, and its differentially expressed profile in Large White and Chinese Erhualian PMSG-hCG stimulated preovulatory ovaries. By comparative sequencing of miR-27a gene in Large White and Chinese Meishan pigs, one SNP (T/C) which created an additional HpaII site was detected. Then associations of this SNP with litter size traits were assessed in Large White (n=142) and DIV (n=140) pig populations. The statistical analysis demonstrated that AA differed from AB (P<0.01) and BB (P<0.05) for total number of piglets born in the first parities, and also differed from AB (P<0.01) for the number of piglets born alive in all parities (P<0.05) in DIV pigs. No significant difference was observed between different genotypes in Large White pigs.


Subject(s)
Genetic Association Studies , Litter Size/genetics , MicroRNAs/genetics , Polymorphism, Single Nucleotide/genetics , Sus scrofa/genetics , Animals , China , Gene Expression Profiling , Gene Frequency/genetics , Genetics, Population , Genotype , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Quantitative Trait, Heritable
18.
Meat Sci ; 87(3): 299-303, 2011 Mar.
Article in English | MEDLINE | ID: mdl-21093991

ABSTRACT

Bone morphogenetic protein 5 (BMP5) gene was suggested to be a potential positional candidate gene for fatness trait QTLs on SSC7. Here by comparative sequencing of BMP5 gene in Meishan (MS) and Large White (LW) pigs, one SNP *131C>T in 3' un-translated region was detected. Association analysis results in 322 LW×MS F(2) pigs showed that CC pigs had thinner back fatness and heavier ham than TC (P<0.05 or P<0.1), and had a higher fat percentage and a lower lean meat percentage (P<0.1) than TT. Moreover, this C/T transition was predicted to alter BMP5 interaction with let-7c and miR-184 by using RNA22 and RNAhybrid. The negative expression of BMP5 gene with let-7c and miR-184 detected from miRNAs overexpression in swine fibroblast, indicating these 2 miRNAs might participate in the translational inhibition of BMP5 gene. Overall, SNP *131C>T might be a good marker for fatness traits.


Subject(s)
Body Composition/genetics , Bone Morphogenetic Protein 5/genetics , MicroRNAs/metabolism , Polymorphism, Single Nucleotide , Sus scrofa/growth & development , Sus scrofa/genetics , 3' Untranslated Regions , Adiposity/genetics , Algorithms , Animals , Binding Sites , Bone Morphogenetic Protein 5/metabolism , Cells, Cultured , Female , Fibroblasts , Gene Frequency , Genetic Association Studies/veterinary , Genetic Markers , Male , Models, Chemical , Mutation , Quantitative Trait Loci/genetics
19.
Clin Exp Immunol ; 161(2): 357-63, 2010 Aug.
Article in English | MEDLINE | ID: mdl-20456412

ABSTRACT

Allogeneic pancreatic islet transplantation theoretically represents a cure for type 1 diabetes. However, current immune suppressive therapies are often associated with undesired side effects. Given this problem, and the shortage of human islet donors, the majority of type 1 diabetes patients cannot currently be offered an islet transplant. However, it has been found that mesenchymal stem cells (MSCs) could exert unique immunosuppressive effects both in vitro and in vivo. Herein we transplanted allogeneic 200 islets alone or in combination with MSCs (3 x 10(6) cells) under the kidney capsules of diabetic C57LB/6 mouse. We found that the ratios of T helper type 1 (Th1) to Th2 and Tc1 to Tc2 were reduced, and the numbers of naive and memory T cells were down-regulated in peripheral blood after transplantation. In addition, the maturation, endocytosis and interleukin-12 secretion of dendritic cell (DCs)-derived bone marrow cells (BMCs) from receptor mice were suppressed. Rejection reaction was alleviated by MSCs which exerted suppressive effects through T lymphocyte subsets and DCs.


Subject(s)
Dendritic Cells/immunology , Diabetes Mellitus, Experimental/therapy , Immunomodulation/immunology , Islets of Langerhans Transplantation/methods , Mesenchymal Stem Cell Transplantation/methods , T-Lymphocytes/immunology , Animals , Antibodies, Monoclonal/immunology , Antibodies, Monoclonal/therapeutic use , Antigens, CD/metabolism , B7-2 Antigen/metabolism , Blood Glucose/metabolism , Bone Marrow Cells/cytology , CD11c Antigen/metabolism , Cell Differentiation/immunology , Dendritic Cells/cytology , Dendritic Cells/metabolism , Dextrans/immunology , Diabetes Mellitus, Experimental/blood , Diabetes Mellitus, Experimental/pathology , Immunoglobulins/metabolism , Interleukin-12/metabolism , Kidney/pathology , Leukocyte Common Antigens/immunology , Lymphocyte Count , Male , Membrane Glycoproteins/metabolism , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Phagocytosis/immunology , T-Lymphocytes/cytology , T-Lymphocytes, Cytotoxic/cytology , T-Lymphocytes, Cytotoxic/immunology , Th1 Cells/cytology , Th1 Cells/immunology , Th2 Cells/cytology , Th2 Cells/immunology , CD83 Antigen
20.
J Appl Genet ; 51(1): 51-7, 2010.
Article in English | MEDLINE | ID: mdl-20145300

ABSTRACT

The contractile protein troponin I (TnI), a constituent protein of the troponin complex located on the thin filaments of striated muscle, is involved in inhibition of calcium-induced myosin ATPase activity (and thus contraction). TnI-slow (slow-twitch skeletal muscle isoform, named TNNI1) and TnI-fast (fast-twitch skeletal muscle isoform, named TNNI2) are muscle-fiber-type-specific proteins, and expression of their genes may affect the composition of muscle fiber, thereby influencing the meat quality traits. Thus, the TnI genes are potential candidate genes for traits related to meat quality in animals. Association of 2 SNPs (EU743939:g.5174T>C in intron 4, and EU743939:g.8350C>A in intron 7) of the TNNI1 gene and a SNP (EU696779:g.1167C>T in intron 3) of the TNNI2 gene with 11 meat quality traits were studied on 334 Large White x Meishan F(2) pigs. In the TNNI1 gene, g.5174T>C and g.8350C>A were found to be significantly associated with intramuscular fat content and meat color value of biceps femoris. The g.5174T>C also showed significant effects on meat color value and marbling score of longissimus dorsi, as well as pH of longissimus dorsi and semispinalis capitis. The g.1167C>T polymorphism in the TNNI2 gene affected significantly the pH of longissimus dorsi, meat color value of longissimus dorsi and semispinalis capitis, marbling score of longissimus dorsi, and intramuscular fat.


Subject(s)
Meat/standards , Muscle, Skeletal/physiology , Polymorphism, Single Nucleotide/genetics , Quantitative Trait, Heritable , Swine/genetics , Troponin I/genetics , Animals , Genotype , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Swine/growth & development
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