1.Association study of schizophrenia and phosphodiesterase 4B gene polymorphism
Li ZHANG ; Yuantang CHEN ; Changjiang HE ; Lei AO ; Yangling XING
Chinese Journal of Behavioral Medicine and Brain Science 2010;19(5):429-431
Objective To detect the genetic association between schizophrenia and polymorphism of phosphodiesterase 4B (PDE4B) gene. Methods Observed in a sample of 98 parent/offspring trios where the proband net the Amerecan Classification and diagnostic Criteria for Mental Disorders The Forth Revised Edition,criteria for schizophrenia using correlation analysis and haplotype relative risk analysis. The polymorphism of phosphodiesterase 4B gene was detected with PCR methods and SNP typing in all nucleus families. Results The rs2180335 al-lele was connected with schizophrenia (P = 0.02131). Allele A was protective factor (Z = -2. 184) and allele G was the hazard factor (Z = 2.184). The frequency of rs2180335 allele A was 0.452 and the allele G was 0.548. The rs1040716, rs 3767311 and rs472952 allele was independence with schizophrenia. Five kinds haplotypes of A/A in the rs2180335-rs3767311, A/A in the rs 3767311-rs472952, A/A/A in the rs2180335-rs3767311-rs472952,A/G/G/A and T/A/A/A in the rs1040716-rs2180335-rs3767311-rs472952 associated with schizophrenia (P values were 0. 028715,0. 034845,0. 024177,0. 023967,0. 010839,genotype frequencies were 0. 223, 0.223,0.127,0.081,0.073). Conclusion It shows an association between schizophrenia and the rs2180335 polymorphism at nucleotide of phosphodiesterase 4B gene in Chinese.
2.Association study between schizophrenia and polymorphism of phosphoserine aminotransferase 1gene
Juan GUO ; Yuantang CHEN ; Changjiang HE ; Li ZHANG ; Yu WU ; Yangling XING ; Lei AO ; Xiang CHEN
Chinese Journal of Behavioral Medicine and Brain Science 2011;20(11):1019-1021
ObjectiveTo detect the association between schizophrenia and polymorphism of phosphoserine aminotransferase 1 ( PSAT1 ) gene.MethodsThe study group included 158 patients with schizophrenia from Xi' an Mental Health Center and the control group included 316 parents.The polymorphism of rs69287125,rs137824326 of phosphoserine aminotransferase 1 gene was detected with PCR methods and SNP typing in all nucleus families by correlation analysis and haplotype relative risk analysis.ResultsThe rs69287125 allele was associated with schizophrenia (P=0.011 ),the G allele was protective factor (Z =-2.31 ) and the A allele was hazarding factor (Z =2.31 ).The rs137824326 allele was associated with schizophrenia (P=0.007 ),the G allele was protective factor ( Z =- 2.54) and the A allele was the hazarding factor( Z =2.54).The haplotypes of A/A and G/G in the rs69287125-rs137824326 were associated with schizophrenia (P =0.021,0.015,Z =2.16,- 1.85).ConclusionThe polymorphism of phosphoserine aminotransferase 1 gene is associated with schizophrenia in Chinese.
3.Association study of schizophrenia and ankyrin repeat and kinase domain containing 1 gene polymorphism
Juan GUO ; Yuantang CHEN ; Changjiang HE ; Li ZHANG ; Yu WU ; Yangling XING ; Lei AO
Chinese Journal of Behavioral Medicine and Brain Science 2010;19(10):912-914
Objective To detect the genetic association between schizophrenia and polymorphism of Ankyrin repeat and kinase domain containing 1 ( ANKK1 ) gene. Methods Observed in a sample of 112 parent/offspring trios where the proband net the American Classification and diagnostic Criteria for Mental Disorders The Forth Revised Edition, criteria for schizophrenia using correlation analysis and haplotype relative risk analysis. The polymorphism of Ankyrin repeat and kinase domain containing 1 gene was detected with PCR methods and SNP typing in all nucleus families. Results The rs2734849 allele was connected with schizophrenia(P= 0. 026). Allele T was protective factor( Z= -2.19) and allele A was the hazard factor( Z=2. 19). The rs4938015,rs7118900 and rs1800497 allele were independence with schizophrenia. Three kinds haplotypes of G/A in the rs7118900 -rs2734849, A/C in the rs2734849 -rs1800497, G/A/C in the rs7118900 -rs2734849 -rs1800497 were associated with schizophrenia ( The P values were 0.032,0. 041,0.046, the genotype frequencies were 0. 36,0.29,0. 17 ).Conclusion It shows an association between schizophrenia and the polymorphism at nucleotide of ankyrin repeat and kinase domain containing 1 gene in Chinese.
4.Regulation effects of intracellular and extracellular Ca2+ on biosynthesis of rosmarinic acid induced by salicylic acid in young seedlings of Salvia miltiorrhiza.
Rong-Rong CAO ; Xiao-Lin DANG ; Bing-Yu XING ; Jing-Yi ZHANG ; Juan-E DONG
China Journal of Chinese Materia Medica 2013;38(20):3424-3431
OBJECTIVETo investigate the effect of intracellular and extracellular Ca2+ on the biosynthesis of rosmarinic acid (RA) induced by salicylic acid in young seedlings of Salvia miltiorrhiza.
METHODYoung seedlings of S. miltiorrhiza were used to select an optimal concentration of salicylic acid (SA), and then use the optimal concentration of SA to investigate the effects of extracellular Ca2+ channel inhibitors Verapamil, LaCl3, intracelluar calmodulin antagonist TFP and intracelluar Ca2+ channel inhibitors LiCl on the biosynthesis of RA and related enzymes.
RESULTSA increased the accumulation of RA and the activities of PAL and TAT, especially the SA of 2 mmol x L(-1) after 24 h. SA improved the accumulation of RA to (40.51 +/- 2.16) mg x g(-1), which was 1.97 times than that of control, and the activities of PAL, TAT were 1.42 times and 1.29 times than those of the control. However, Vp, LaCl3, TFP, LiCl inhibited the effects of SA evidently.
CONCLUSIONCa2+ plays a key role in the regulation of the induction process.
Calcium ; metabolism ; Cinnamates ; metabolism ; Depsides ; metabolism ; Gene Expression Regulation, Plant ; Plant Proteins ; genetics ; metabolism ; Salicylic Acid ; metabolism ; Salvia miltiorrhiza ; genetics ; growth & development ; metabolism ; Seedlings ; genetics ; growth & development ; metabolism
5.Cryptosporidium suis Infection in Post-Weaned and Adult Pigs in Shaanxi Province, Northwestern China.
Qing LIN ; Xing Ye WANG ; Jian Wen CHEN ; Ling DING ; Guang Hui ZHAO
The Korean Journal of Parasitology 2015;53(1):113-117
Cryptosporidium spp., ubiquitous enteric parasitic protozoa of vertebrates, recently emerged as an important cause of economic loss and zoonosis. The present study aimed to determine the distribution and species of Cryptosporidium in post-weaned and adult pigs in Shaanxi province, northwestern China. A total of 1,337 fresh fecal samples of post-weaned and adult pigs were collected by sterile disposable gloves from 8 areas of Shaanxi province. The samples were examined by Sheather's sugar flotation technique and microscopy atx400 magnification for Cryptosporidium infection, and the species in positive samples was further identified by PCR amplification of the small subunit (SSU) rRNA gene. A total of 44 fecal samples were successfully amplified by the nested PCR of the partial SSU rRNA, with overall prevalence of 3.3%. The average prevalence of Cryptosporidium infection in each pig farms ranged from 0 to 14.4%. Species identification by sequencing of SSU rRNA gene revealed that 42 (3.1%) samples were Cryptosporidium suis and 2 (0.15%) were Cryptosporidium scrofarum. C. suis had the highest prevalence (7.5%) in growers and the lowest in breeding pigs (0.97%). C. suis was the predominant species in pre-weaned and adult pigs, while C. scrofarum infected pigs older than 3 months only. A season-related difference of C. suis was observed in this study, with the highest prevalence in autumn (5.5%) and the lowest (1.7%) in winter. The present study provided basic information for control of Cryptosporidium infection in pigs and assessment of zoonotic transmission of pigs in Shaanxi province, China.
Animals
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China/epidemiology
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Cluster Analysis
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Cryptosporidiosis/*epidemiology/*parasitology
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Cryptosporidium/classification/genetics/*isolation & purification
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DNA, Protozoan/chemistry/genetics
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DNA, Ribosomal/chemistry/genetics
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Feces/parasitology
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Molecular Sequence Data
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Phylogeny
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Polymerase Chain Reaction
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Prevalence
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RNA, Ribosomal, 18S/genetics
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Seasons
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Sequence Analysis, DNA
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Swine
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Swine Diseases/*epidemiology/*parasitology
6.Cryptosporidium suis Infection in Post-Weaned and Adult Pigs in Shaanxi Province, Northwestern China.
Qing LIN ; Xing Ye WANG ; Jian Wen CHEN ; Ling DING ; Guang Hui ZHAO
The Korean Journal of Parasitology 2015;53(1):113-117
Cryptosporidium spp., ubiquitous enteric parasitic protozoa of vertebrates, recently emerged as an important cause of economic loss and zoonosis. The present study aimed to determine the distribution and species of Cryptosporidium in post-weaned and adult pigs in Shaanxi province, northwestern China. A total of 1,337 fresh fecal samples of post-weaned and adult pigs were collected by sterile disposable gloves from 8 areas of Shaanxi province. The samples were examined by Sheather's sugar flotation technique and microscopy atx400 magnification for Cryptosporidium infection, and the species in positive samples was further identified by PCR amplification of the small subunit (SSU) rRNA gene. A total of 44 fecal samples were successfully amplified by the nested PCR of the partial SSU rRNA, with overall prevalence of 3.3%. The average prevalence of Cryptosporidium infection in each pig farms ranged from 0 to 14.4%. Species identification by sequencing of SSU rRNA gene revealed that 42 (3.1%) samples were Cryptosporidium suis and 2 (0.15%) were Cryptosporidium scrofarum. C. suis had the highest prevalence (7.5%) in growers and the lowest in breeding pigs (0.97%). C. suis was the predominant species in pre-weaned and adult pigs, while C. scrofarum infected pigs older than 3 months only. A season-related difference of C. suis was observed in this study, with the highest prevalence in autumn (5.5%) and the lowest (1.7%) in winter. The present study provided basic information for control of Cryptosporidium infection in pigs and assessment of zoonotic transmission of pigs in Shaanxi province, China.
Animals
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China/epidemiology
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Cluster Analysis
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Cryptosporidiosis/*epidemiology/*parasitology
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Cryptosporidium/classification/genetics/*isolation & purification
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DNA, Protozoan/chemistry/genetics
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DNA, Ribosomal/chemistry/genetics
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Feces/parasitology
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Molecular Sequence Data
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Phylogeny
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Polymerase Chain Reaction
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Prevalence
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RNA, Ribosomal, 18S/genetics
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Seasons
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Sequence Analysis, DNA
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Swine
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Swine Diseases/*epidemiology/*parasitology
7.Effects of calcium on synthesis of rosmarinic acid and related enzymes in suspension cultures of Salvia miltiorrhiza.
Liancheng LIU ; Juan'e DONG ; Jingyi ZHANG ; Xiaolin DANG ; Bingyu XING ; Xiling YANG
Chinese Journal of Biotechnology 2012;28(11):1359-1369
We studied the influence of the concentration of Ca2+ (0-50 mmol/L) in culture medium on the synthesis of rosmarinic acid (RA) and related enzymes in Salvia miltiorrhiza suspension cultures. Using verpamil (VP, a calcium channel antagonist) and ionophore A23187, we studied the mechanism of secondary metabolites of Salvia miltiorrhiza suspension cultures influenced by the concentration of Ca2+ in the culture medium. The synthesis of intracellular RA in 6-day incubation was significantly dependent on the medium Ca2+ concentration. At the optimal Ca2+ concentration of 10 mmol/L, a maximal RA content of 20.149 mg/g biomass dry weight was reached, which was about 37.3% and 20.4% higher than that at Ca2+ concentrations of 1 and 3 mmol/L, respectively. The variation of the activity of PAL and TAT, two key enzymes of the two branches of RA, could be affected by the concentration of Ca2+ in culture medium. The change of their activity occurred prior to the accumulation of RA, which suggested both of the key enzymes be involved in the synthesis of RA. Meanwhile, the enzymatic action of PAL was more distinct than TAT. The treatment of VP and A23187, respectively, indicated that the influence of RA affected by the concentration of Ca2+ in the culture medium was accomplished by the intracellular Ca2+, and the flow of Ca2+ from the extracellular to the intracellular environment could also participate in this process.
Calcium
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pharmacology
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Cinnamates
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metabolism
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Culture Media
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Culture Techniques
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methods
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Depsides
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metabolism
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Phenylalanine Ammonia-Lyase
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metabolism
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Salvia miltiorrhiza
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chemistry
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enzymology
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growth & development
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Tyrosine Transaminase
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metabolism
8.Optimization of bacterial cellulose fermentation medium and observation of bacterial cellulose ultra-micro-structure.
Ruiqin WU ; Shuangkui DU ; Zhixi LI ; Xiaohui XING ; Dongyan SHAO ; Yanli FAN ; Bo LI ; Xueliang ZHANG ; Lijun BU
Chinese Journal of Biotechnology 2008;24(6):1068-1074
In order to improve the yield of bacterial cellulose (BC), the fermentation medium of BC-producing strain J2 (Gluconobacter) was optimized, and BC ultra-micro-structure was observed. Initially, Plackett-Burman design was employed to evaluate eight variables which were relevant to BC production. Three statistically significant parameters including yeast extract, ZnSO4, ethanol were selected and other 5 variables were not significant (P > 0.05). The optimized levels of three variables were defined by Box-Behnken design and response surface methodology (RSM). BC ultra-micro-structure was observed by scanning electron microscope (SEM) with cotton cellulose as comparison. The results indicated that the BC yield under the optimum fermentation medium was 11.52 g/100 mL, which was as 1.35 times as that under the original fermentation medium. The SEM photos manifested that bacterial cellulose ribbon, with a diameter less than 0.1 microm, was less than cotton cellulose ribbon. The bacteria inside the cellulose net were eliminated after the NaOH treatment.
Cell Culture Techniques
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Cellulose
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biosynthesis
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ultrastructure
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Culture Media
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chemistry
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Fermentation
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Gluconobacter
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cytology
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metabolism
9.The application of co-culture system on the in vitro development of bovine somatic nuclear transferred embryos.
Yue-Hong WU ; Zhi-Xing AN ; Yong ZHANG ; Xiang-Chen LI ; Yu-Long HE ; Feng-Jun LIU
Chinese Journal of Biotechnology 2006;22(2):306-310
To establish a co-culture system of nuclear transferred embryos in bovine, effects of co-culture cell types, passages and cryopreservation as well as addition of BFF or FBS were investigated. The results showed that embryos co-cultured with oviductal epithelial cell and granulosa cell achieved significantly higher blastocyst rate compared with the control group (P < 0.05) and co-cultured with oviductal epithelial cell had more embryo cell number than those with granulosa cell. Passages of co-culture cells significantly affected the blastocyst rate and embryo cell number (P < 0.05), and cryopreservation decreased the blastocyst rate and embryo cell number remarkably. Supplemention of BFF increased blastocyste rate significantly (P < 0.05). In conclusion, co-cultured with fresh primary oviductal epithelial cell along with addition of 10% BFF in SOFaa could improve development of nuclear transferred bovine embryo in vitro.
Animals
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Cattle
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Cellular Reprogramming
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Cloning, Organism
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methods
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Coculture Techniques
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Culture Media
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Embryo, Mammalian
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cytology
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drug effects
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Embryonic Development
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Epithelial Cells
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cytology
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drug effects
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Fallopian Tubes
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cytology
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Female
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Granulosa Cells
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cytology
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drug effects
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Nuclear Transfer Techniques
10.Preparation and identification of recombinant adenoviruses carrying short hairpin RNA targeting parathyroid hormone related protein of goat.
Ruifang XING ; Huiling ZHENG ; Xuemei LIU ; Linhui YAN ; Junhui AN ; Zhenyu YANG ; Zhenzhen ZHU
Chinese Journal of Biotechnology 2011;27(11):1555-1563
Parathyroid hormone related protein (PTHrP) has important biological functions in calcium metabolism. The aim of this study was to silence the expression of PTHrP by RNA interference and recombinant adenovirus, and to provide a material to investigate the relative functions of PTHrP in goat mammary gland epithelial cell. The Block-iT shRNA interference system was used in this experiment. We designed and synthesized two pairs of complementary single-strand DNA oligonucleotides (shRNA-322/357) targeting two different sites of PTHrP mRNA. Then the oligonucleotides were inserted into shuttle vector pENTR/CMV-GFP/U6. After detection of the interference efficiency by Western blotting, we chose pENTR/CMV-GFP/U6-322 and adenovirus backbone vector pAD/PL-DEST to produce recombinant vector pAD/PL-DEST/CMV-GFP/U6-322. The first generation recombinant adenovirus particles (AD-PTHrP-322) were produced and further amplified by transfecting HEK-293 cells. The titer of the recombinant adenovirus reached 2.0 x 1(9) PFU/mL determined by TCID50 assays. The result of real-time quantitative PCR indicated that mRNA expression levels of gene were reduced 29.2%, 68.1% and 82.6% (P < 0.05), respectively, when goat mammary gland epithelial cells were infected with AD-PTHrP-322 after 24, 48 and 72 h, in which PTHrP. Western blotting also showed that the expression of PTHrP was reduced by infecting the cells with AD-PTHrP-322. AD-PTHrP-322 has been proved with significant interference effect on expression of PTHrP.
Adenoviridae
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genetics
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metabolism
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Animals
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Epithelial Cells
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metabolism
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Female
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Genetic Vectors
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genetics
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Goats
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HEK293 Cells
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Humans
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Mammary Glands, Animal
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cytology
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Parathyroid Hormone-Related Protein
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biosynthesis
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genetics
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RNA Interference
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RNA, Messenger
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biosynthesis
;
genetics
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RNA, Small Interfering
;
genetics
;
Recombinant Proteins
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biosynthesis
;
genetics
;
isolation & purification
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Transfection