AIM: To study gastric mucosal interleukine-8 (IL-8) mRNA expression, the cytotoxin-associated gene A (cagA) mutation, and serum pepsinogen (PG)?I/II ratio related risk in Thai gastric cancer.METHODS: There were consen...AIM: To study gastric mucosal interleukine-8 (IL-8) mRNA expression, the cytotoxin-associated gene A (cagA) mutation, and serum pepsinogen (PG)?I/II ratio related risk in Thai gastric cancer.METHODS: There were consent 134 Thai non-cancer volunteers who underwent endoscopic narrow band imaging examination, and 86 Thais advance gastric cancer patients who underwent endoscopic mucosal biopsies and gastric surgery. Tissue samples were taken by endoscopy with 3 points biopsies. The serum PG?I, II, and Helicobacter pylori (H. pylori) immunoglobulin G (IgG) antibody for H. pylori were tested by enzyme-linked immunosorbent assay technique. The histopathology description of gastric cancer and non-cancer with H. pylori detection was defined with modified Sydney Score System. Gastric mucosal tissue H. pylori DNA was extracted and genotyped for cagA mutation. Tissue IL-8 and cyclooxygenase-2 (COX-2) mRNA expression were conducted by real time relative quantitation polymerase chain reaction. From 17 Japanese advance gastric cancer and 12 benign gastric tissue samples, all were tested for genetic expression with same methods as well as Thai gastric mucosal tissue samples. The multivariate analysis was used for the risk study. Correlation and standardized t-test were done for quantitative data, P value < 0.05 was considered as a statistically significant.RESULTS: There is a high non cagA gene of 86.8 per cent in Thai gastric cancer although there are high yields of the East Asian type in the positive cagA. The H. pylori infection prevalence in this study is reported by combined histopathology and H. pylori IgG antibody test with 77.1% and 97.4% of sensitivity and specificity, respectively. The serum PG?I/II ratio in gastric cancer is significantly lower than in the non-cancer group, P = 0.045. The serum PG?I/II ratio of less than 3.0 and IL-8 mRNA expression ≥ 100 or log10 ≥ 2 are significant cut off risk differences between Thai cancer and non-cancer, P = 0.03 and P < 0.001, respectively. There is a significantly lower PGI/II ratio in Japanese than that in Thai gastric cancer, P = 0.026. Serum PG?I/II ratio at cut off less than 3.0 and IL-8 mRNA expression Raw RQ > 100 or log10 > 2 are significantly difference between Thai cancer group when compared to non-cancer group, P = 0.013 and P < 0.001, respectively. In the correlation study, low PG?I/II ratio does not associate with chronic atrophic gastritis severity score in Thais non-cancer cases. However, there is a trend, but not significant convert correlation between IL-8 mRNA expression level and low PG?I/II ratio in Thai positive H. pylori infection. The high expression of IL-8 gene demonstrates a poorer prognosis by stage and histology.CONCLUSION:Predominant gastric mucosal IL-8 mRNA expression level, H. pylori infection, and low PG?I/II ratio are relative risks for Thai gastric cancer without correlation with cagA mutation.展开更多
AIM： To investigate the expression of gastrin-releasing peptide （GRP） and GRP-receptor mRNA in non-tumor tissues of the human esophagus, gastrointestinal tract, pancreas and gallbladder using molecular biology tech...AIM： To investigate the expression of gastrin-releasing peptide （GRP） and GRP-receptor mRNA in non-tumor tissues of the human esophagus, gastrointestinal tract, pancreas and gallbladder using molecular biology techniques. METHODS： Poly A^＋ mRNA was isolated from total RNA extracts using an automated nucleic acid extractor and, subsequently, converted into single-stranded cDNA （sscDNA）. PCR amplifications were carried out using genespecific GRP and GRP-receptor primers. The specificity of the PCR amplicons was further confirmed by Southern blot analyses using gene-specific GRP and GRP-receptor hybridization probes. RESULTS： Expression of GRP and GRP-receptor mRNA was detected at various levels in nearly all segments of the non-tumor specimens analysed, except the gallbladder. In most of the biopsy specimens, coexpression of both GRP and GRP-receptor mRNA appeared to take place. However, expression of GRP mRNA was more prominent than was GRP-receptor mRNA. CONCLUSION： GRP and GRP-receptor mRNAs are expressed throughout the gastrointestinal tract and provides information for the future mapping and determination of its physiological importance in normal and tumor cells.展开更多
Mammalian macrophage migration inhibitory factor （MIF） plays an important role as an indispensable mediator in the pathogenesis of inflammatory disease like septicemia, but little is known about the role of MIF homo...Mammalian macrophage migration inhibitory factor （MIF） plays an important role as an indispensable mediator in the pathogenesis of inflammatory disease like septicemia, but little is known about the role of MIF homologue in fish septicemia. The authors have cloned the MIF homologue in large yellow croaker Pseudosciaena crocea （LycMIF） using RACE approach. The full-length cDNA of LycMIF was 634 bases and contained an ORF of 345 bases encoding a protein of 115 amino acid residues. As demonstrated by RT-PCR and QRT-PCR assay, MIF mRNAs were constitutively expressed in 11 selected tissues and were abundant in brain and liver. Moreover, the LycMIF transcripts in the liver and head kidney were responsive to bacteria infection and could be significantly up-regulated. Our results provide the first direct evidence that fish MIF was implicated in pathogenesis of fish vibrosis and play an important role in response to bacteria infection.展开更多
Previous studies on mammals showed that peroxisome proliferator-activated receptor gamma coactivator-1α(PGC-1α)played a prominent role in regulating muscle fiber type transition and composition.However,the role of P...Previous studies on mammals showed that peroxisome proliferator-activated receptor gamma coactivator-1α(PGC-1α)played a prominent role in regulating muscle fiber type transition and composition.However,the role of PGC-1αin chicken muscle has seldom been explored.To investigate the effect of PGC-1αon chicken skeletal muscles in this study,the PGC-1αgene was overexpressed or silenced in chicken primary myoblasts by using lentivirus,and then the effects of the PGC-1αgene overexpression and knockdown on the mRNA expression profile of genes related to myofiber type specificity were examined during fiber formation.The results showed that overexpression of PGC-1αfrom proliferation to differentiation was accompanied by the up-regulated expression of Pax7,MyoD,and CnAα,which was significantly(P<0.01)increased after one day of transfection(1 I).The enhancement of MyoG,MEF2 c,and MyHC SM expression lagged,which was improved significantly(P<0.01)after four days of transfection(1 I3 D).Overexpression of PGC-1αdecreased(P<0.01)the MyHC FWM expression after four days of transfection(1 I3 D),and it had no significant impact(P>0.05)on the expression of CnB1,NFATc3,and MyHC FRM during myofiber formation.The effective silence(P<0.01)of PGC-1αby lentivirus mediating short hairpin RNA(shRNA)was detected after four days of transfection(1 I3 D)in cultures,and the lack of its function in chicken primary myoblasts significantly(P<0.01)down-regulated the expression of Pax7,MyoD,CnAα,MyoG,MEF2 c,and MyHC SM,significantly(P<0.01)up-regulated the expression of MyHC FWM,and had no significant impact(P>0.05)on the expression of CnB1,NFATc3,and MyHC FRM.These results indicated that the role of PGC-1αin regulating the fiber type specificity of chicken skeletal muscles might be similar to that in mammals,which interplayed with key genes related to myocyte differentiation and calcineurin signaling pathway.展开更多
The H19 gene, which is imprinted with preferential expression from the maternal allele, was one of the first identified imprinting genes in mammals. Recent studies revealed that correct imprinting of the H19 gene play...The H19 gene, which is imprinted with preferential expression from the maternal allele, was one of the first identified imprinting genes in mammals. Recent studies revealed that correct imprinting of the H19 gene plays a vital role in human spermatogenesis. To investigate whether imprinting defects were associated with the hybrid sterility of male cattle-yak, the methylation patterns of the H19 imprinting control region （ICR） and H19 mRNA expression in the testes of cattle-yak, yak, and cattle were examined. The results showed that the 3rd CCCTC-binding factor （CTCF） site of the H19 ICR was significantly hypomethylated in the testes of cattle-yak compared with yak or cattle. As expected, H19 was expressed at a significantly higher level in cattle-yak than in yak or cattle. These results suggest that imprinting defects of the CTCF- binding site in the HI9 ICR were possibly associated with disturbed spermatogenesis of male cattle-yak. Thus, we propose that disorders in H19 imprinting, resulting in an increased H19 mRNA expression, might contribute to the sterility of F1 male hybrids between cattle and yak.展开更多
A homologue of the lower vertebrates translationally controlled tumor protein (TCTP) was cloned from the marine fish Japanese sea perch (Lateolabrax japonicus) by the technology of homology cloning. The full-length cD...A homologue of the lower vertebrates translationally controlled tumor protein (TCTP) was cloned from the marine fish Japanese sea perch (Lateolabrax japonicus) by the technology of homology cloning. The full-length cDNA sequence of the sea perch TCTP gene contained a 5' untranslated region (UTR) of 47 bp, a 3' UTR of 433 bp, and a putative open reading frame (ORF) of 510 bp encoding a polypeptide of 170 amino acids. The deduced amino acid sequence of the sea perch TCTP gene showed a high similarity to that of zebrafish, rohu, rabbit, chicken and human. Sequence analysis revealed there were a signature sequence of TCTP family, an N-glycosylation site, and five Casein kinase phosphorylation sites in the sea perch TCTP. The temporal expression of TCTP genes in healthy and lipopolysaccharide (LPS) challenged fishes was measured by semi-quantitative reverse transcription-PCR (RT-PCR). The results indicated that LPS could up-regulate the expression of sea perch TCTP in the examined tissues, including head-kidney, spleen and liver.展开更多
Background:Maternal nutrition during gestation affects fetal development,which has long-term programming effects on offspring postnatal growth performance.With a critical role in protein and lipid metabolism,essential...Background:Maternal nutrition during gestation affects fetal development,which has long-term programming effects on offspring postnatal growth performance.With a critical role in protein and lipid metabolism,essential fatty acids can influence the development of muscle and adipose tissue.The experiment investigated the effects of late gestation supplements(77 d prepartum),either rich in saturated and monounsaturated fatty acids(CON;155 g/cow/d EnerGII)or polyunsaturated fatty acids(PUFA;80 g/cow/d Strata and 80 g/cow/d Prequel),on cow performance and subsequent calf growth performance as well as mRNA expression in longissimus muscle(LM)and subcutaneous adipose tissue at birth and weaning.Results:There was no difference(P≥0.34)in cow body weight(BW)or body condition score from presupplementation through weaning.Relative concentrations of C18:3n-3 and C20:4n-6 decreased(P≤0.05)to a greater extent from mid-supplementation to calving for PUFA compared with CON cows.Cow plasma C20:0,C20:5n-3,and C22:6n-3 were increased(P≤0.01)in PUFA during supplementation period.At birth,PUFA steers had greater(P=0.01)plasma C20:5n-3.No differences(P≥0.33)were detected in steer birth BW or dam milk production,however,CON steers tended(P=0.06)to have greater pre-weaning average daily gain and had greater(P=0.05)weaning BW compared with PUFA.For mRNA expression in steers:MYH7 and C/EBPβin LM increased(P≤0.04)to a greater extent from birth to weaning for PUFA compared with CON;MYF5 in LM and C/EBPβin adipose tissue tended(P≤0.08)to decrease more from birth to weaning for CON compared with PUFA;SCD in PUFA adipose tissue tended(P=0.08)to decrease to a greater extent from birth to weaning than CON.In addition,maternal PUFA supplementation tended(P=0.08)to decrease MYOG mRNA expression in LM and decreased(P=0.02)ZFP423 in adipose tissue during the pre-weaning stage.Conclusions:Late gestation PUFA supplementation decreased pre-weaning growth performance of the subsequent steer progeny compared with CON supplementation,which could have been a result of downregulated mRNA expression of myogenic genes during pre-weaning period.展开更多
The effects of dietary lysine on production performance,serum concentrations of metabolites,growth hormone （GH）,insulin-like growth factor-I （IGF-I） and IGF-I mRNA expression in growing rabbits were examined.One h...The effects of dietary lysine on production performance,serum concentrations of metabolites,growth hormone （GH）,insulin-like growth factor-I （IGF-I） and IGF-I mRNA expression in growing rabbits were examined.One hundred weaned New Zealand rabbits were allocated to individual cages and randomly offered a diet containing 5.5 （L1）,6.5 （L2）,7.5 （L3）,8.5 （L4）,or 9.5 g （L5） lysine per kg diet.The results showed that the average daily gain （ADG） of the rabbits from L3,L4 or L5 was higher than those from L1 or L2 （P 〈 0.05）.The feed gain ratio （F/G） in the rabbits from L4 or L5 was lower than those from L1 or L2 （P 〈 0.05）.Dietary lysine did not affect serum concentrations of total protein （TP）,glucose,insulin （INS）,and growth hormone （GH） （P 〉 0.05）.The quadratic effects of lysine on the serum urea nitrogen （SUN） concentration was detected （P = 0.035）.Serum IGF-I concentrations had a trend to increase quadratically with the increasing dietary lysine （P = 0.07）.A significant correlation was found between serum IGF-I concentrations （x,ng mL-1） and ADG （y,g kg-1）： y = -0.017x2 ＋ 1.984x ＋ 20.87 （R2 = 0.8982,P = 0.003）.The relative abundance of hepatic and muscular IGF-I mRNA tended to increase with increasing dietary lysine levels （P = 0.053 and 0.082,respectively）.Providing the diets mainly consisted of corn,wheat bran and peanut vine,the most appropriate dietary lysine level for growing meat rabbits from weaning to 70 d old was found to be 8.5 g kg-1,and IGF-I may be an important factor controlling growth of weaned rabbits.展开更多
AIM： To investigate the effects of arginine （Arg）-enriched diets before sepsis and/or Arg-containing total parenteral nutrition （TPN） after sepsis or both on cytokine mRNA expression levels in splenocytes of rats...AIM： To investigate the effects of arginine （Arg）-enriched diets before sepsis and/or Arg-containing total parenteral nutrition （TPN） after sepsis or both on cytokine mRNA expression levels in splenocytes of rats with gut-derived sepsis. METHODS： Rats were assigned to four experimental groups. Groups 1 and 2 were fed with a semipurified diet, while groups 3 and 4 had part of the casein replaced by Arg which provided 2% of the total calories. After the rats were fed with these diets for 10 d, sepsis was induced by cecal ligation and puncture （CLP）, at the same time an internal jugular vein was cannulated. All rats were maintained on TPN for 3 d. Groups 1 and 3 were infused with conventional TPN, while groups 2 and 4 were supplemented with Arg which provided 2% of the total calories in the TPN solution. All rats were killed 3 d after CLP to examine their splenocyte subpopulation distribution and cytokine expression levels. RESULTS： Plasma interleukin （IL）-2, IL-4, tumor necrosis factor-α （TNF-α） and interferon （IFN-γ） were not detectable 3 d after CL.P. There were no differences in the distributions of CD45Ra＋, CD3＋, CD4＋, and CD8＋ cells in whole blood and splenocytes among the four groups. The splenocyte IL-2 mRNA expression in the Arg-supplemented groups was significantly higher than that in group 1. IL-4 mRNA expression in groups 3 and 4 was significantly higher than that in groups 1 and 2. The mRNA expression of IL-10 and IFN-γ was significantly higher in group 4 than in the other three groups. There was no difference in TNF-α mRNA expression among thefour groups CONCLUSION： The influence of Arg on the whole blood and splenic lymphocyte subpopulation distribution is not obvious. However, Arg administration, especially before and after CLP, significantly enhances the mRNA expression levels of Thl and Th2 cytokines in the spleen of rats with gut-derived sepsis.展开更多
Summary： In adult liver, CYP3A4 plays an important role in the metabolism of a wide range of en- dogenous and exogenous compounds. To investigate whether there is a single nucleotide polymorphism （SNP） of CYP3A4 in...Summary： In adult liver, CYP3A4 plays an important role in the metabolism of a wide range of en- dogenous and exogenous compounds. To investigate whether there is a single nucleotide polymorphism （SNP） of CYP3A4 intron 2 in the liver and its effects on the mRNA expression and enzymatic activity of CYP3A4, genomic DNA was extracted from 96 liver tissue samples obtained from patients who had undergone liver surgery. An SNP of CYP3A4 intron 2 was identified by polymerase chain reaction （PCR）-single-strand confirmation polymorphism and DNA sequencing. The mRNA expression of CYP3A4 was determined by the fluorescence quantitative PCR technique. The enzymatic activity of CYP3A4 was measured using erythromycin and testosterone as probe substrates. Twelve patients were found to have the SNP/T4127G CYP3A4 within intron 2. The mRNA levels of CYP3A4 in wild-type and SNP/T4127G samples were 2.62±1.09 and 2.79±1.63, respectively （P〉0.05）. Erythromycin N-demethylase activity in wild-type and SNP/T4127G samples were 121.2±32.8 and 124.7±61.6 nmol·mg^-1min^-1, respectively （P〉0.05）. The activity of testosterone 613-hydroxylase was significantly different between wild-type （648±173 pmol·mg^-1·min^-1） and SNP/T4127G samples （540-4-196 pmol.mg-l-minl; P〈0.05）. In conclusion, the SNP/T4127G of CYP3A4 intron 2 exists in the liver. This SNP does not affect the mRNA expression of CYP3A4 but significantly decreases the hepatic micro- somal testosterone 613-hydroxylase activity of CYP3A4. Furthermore, this study indicates that the ap- propriate selection of probe substrates is very important in studying the relationship between the geno- type and phenotype of CYP3A4.展开更多
AMP deaminase catalyzes the conversion of AMP into IMP and ammonia. In the present study, a full-length cDNA of AMPD1 from skeletal muscle of Japanese flounder Paralichthys olivaceus was cloned and characterized. The ...AMP deaminase catalyzes the conversion of AMP into IMP and ammonia. In the present study, a full-length cDNA of AMPD1 from skeletal muscle of Japanese flounder Paralichthys olivaceus was cloned and characterized. The 2 526 bp cDNA contains a 5'-UTR of 78 bp, a 3'-UTR of 237 bp and an open reading frame (ORF) of 2 211 bp, which encodes a protein of 736 amino acids. The predicted protein contains a highly conserved AMP deaminase motif (SLSTDDP) and an ATP-binding site sequence (EPLMEEYAIAAQVFK). Phylogenetic analysis showed that the AMPD1 and AMPD3 genes originate from the same branch, but are evolutionarily distant from the AMPD2 gene. RT-PCR showed that the flounder AMPD1 gene was expressed only in skeletal muscle. QRT-PCR analysis revealed a statistically significant 2.54 fold higher level of AMPD1 mRNA in adult muscle (750±40 g) compared with juvenile muscle (7.5±2 g) (P<0.05). HPLC analysis showed that the IMP content in adult muscle (3.35±0.21 mg/g) was also statistically significantly higher than in juvenile muscle (1.08±0.04 mg/g) (P<0.05). There is a direct relationship between the AMPD1 gene expression level and IMP content in the skeletal muscle of juvenile and adult flounders. These results may provide useful information for quality improvement and molecular breeding of aquatic animals.展开更多
BACKGROUND： Somatostatin is abundant in the hypothalamus, cerebral cortex, limbic system, and mesencephalon. Somatostatin mRNA expression in the brain of rats with spleen deficiency is noticeably reduced, as well as ...BACKGROUND： Somatostatin is abundant in the hypothalamus, cerebral cortex, limbic system, and mesencephalon. Somatostatin mRNA expression in the brain of rats with spleen deficiency is noticeably reduced, as well as attenuation of cognitive function. OBJECTIVE： To observe the interventional effect of Guipi decoction on somatostatin level and somatostatin receptor 1 （SSTRl） mRNA expression in different encephalic regions of rats with spleen deficiency, and to compare the interventional effects of Guipi decoction, Chaihu Shugan powder, and Tianwang Buxin pellet. DESIGN： A randomized controlled observation. SETTING： Basic Medical College, Beijing University of Traditional Chinese Medicine. MATERIALS： Fifty adult Wistar male rats, of clean grade, weighing （160 ± 10） g, were provided by Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd. The protocol was performed in accordance with ethical guidelines for the use and care of animals. Somatostatin 1 polyclonal anti-rabbit antibody and SSTRl in situ hybridization kit were provided by Department of Neuroanatomy, Shanghai Second Military Medical University of Chinese PLA. The drug for developing rat models of spleen deficiency was composed of Dahuang, Houpu and Zhishi, and prepared at 2：1：1. Guipi decoction, Chaihu Shugan powder, and Tianwang Buxin pellet recipes were made according to previous studies. METHODS： This study was performed at the Basic Medical College, Beijing University of Traditional Chinese Medicine from March 2002 to March 2005. The rats were randomly divided into 5 groups, with 10 rats in each group： normal, model, Guipi decoction, Chaihu Shugan powd.er, and Tianwang Buxin pellet groups. Rat models of the latter 4 groups were developed by methods of purgation with bitter and cold nature drugs, improper diet, and overstrain. The rats received 7.5 g/kg of the drugs each morning and were fasted every other day, but were allowed free access to water at all times. The rats were forced to swim in 25 ℃ water until fatigued. Rats in the normal group were intragastrically administered the same amount of normal saline. Rats in the Guipi decoction, Chaihu Shugan powder, and Tianwang Buxin pellet groups were intragastrically administered 7.5 g/kg Guipi decoction, Chaihu Shugan powder, and Tianwang Buxin pellet, respectively, every afternoon. All rats were treated for 6 weeks. MAIN OUTCOME MEASURES： Somatostatin protein and SSTRI mRNA expression in the ventral nucleus of hypothalamus, hippocampal CAl region, and cortex of prefrontal lobe were determined by immunohistochemistry and in situ hybridization, respectively. RESULTS： Fifty rats were included in the final analysis. In the model group, expression of somatostatin protein and SSTRl mRNA in the ventral nucleus of hypothalamus, hippocampal CAl region, and cortex of prefrontal lobe were significantly less than in the normal group （P 〈 0.01）. Above-mentioned indices were identical in the Chaihu Shugan powder and model groups. However, expression of somatostatin protein and SSTRl mRNA were significantly higher in the Guipi decoction group compared to model group （P 〈 0.01）. In the Tianwang Buxin pellet group, SSTRl mRNA expression in rat ventral nucleus of hypothalamus and somatostatin level in rat hippocampal CAl region and cortex of prefrontal lobe, as well as ventral nucleus of hypothalamus, were significantly higher compared to model group （P 〈 0.01 ）. CONCLUSION： Somatostatin level and SSTRl mRNA expression in rats with spleen deficiency were lower than in normal rats. Guipi decoction and Tianwang Buxin pellet up-regulated somatostatin level and SSTRl mRNA expression.展开更多
Objective: To look for additional markers of molecular biology response to anastrozole, a new aromatase inhibitor, on the growth and mRNA expression level of MCF-7 cell. Methods: We investigated the effect of anastrzo...Objective: To look for additional markers of molecular biology response to anastrozole, a new aromatase inhibitor, on the growth and mRNA expression level of MCF-7 cell. Methods: We investigated the effect of anastrzole on growth and gene expression in the human breast cancer cell line MCF-7 and compared with the most widely used antiestrogen tamoxifen. We chose 4 genes to examine regulation of gene expression of estrogen regulated genes: PR A, PR B, ErbB-2 and cyclin D1. Results: Compared with the tamoxifen, a statistically significant growth inhibition was observed with anastrozole. The PR A, PR B and cyclin D1 mRNA level in anastrozole treated cells was sigificantly below the level in tamoxifen treated cells (P<0. 05). They had agonistic effect on ErbB gene (P>0. 05). Conclusion: The third generation of aromatase inhibitors anastrozole exert more inhibit function in some expression of estrogen regulated genes than tomoxifen in MCF-7 cell line.展开更多
BACKGROUND: It has been demonstrated that adenosine can induce glial cell to release cytochrome C, enhance expression of apoptotic gene bax, inhibit anti-apoptotic gene bcl-2, and activate caspase-3 to apoptosis; Wher...BACKGROUND: It has been demonstrated that adenosine can induce glial cell to release cytochrome C, enhance expression of apoptotic gene bax, inhibit anti-apoptotic gene bcl-2, and activate caspase-3 to apoptosis; Whereas inosine can inhibit neuronal apoptosis which is similar to bcl-2. OBJECTIVE: To observe the effects of inosine on neuronal apoptosis and expression of cytochrome C mRNA in rats after focal cerebral ischemia/reperfusion, and analyze the pathway of its neuroprotective effect. DESIGN: A randomised controlled animal trial. SETTINGS: Department of Neurology, Rongcheng Second People's Hospital; Department of Neurology, Affiliated Union Hospital, Tongji Medical College, Huazhong University of Science and Technology. MATERIALS: Sixty-eight rats, weighing 230-280 g and clean grade, were used. TdT-mediated dUTP-biotin nick end labeling (TUNEL) and cytochrome C mRNA in situ hybridization kits and DAB staining kit were purchased from Wuhan Boster Biological Co., Ltd.; Inosine injection [200 mg (2 mL) each] from Qingdao First Pharmaceutical Factory. METHODS: The experiment was accomplished in the animal experimental center in Tongji Medical College of Huazhong University of Science and Technology from December 2003 to June 2005. ① Sixty-four rats were made into focal ischemia by middle cerebral artery occlusion (MCAO) with a nylon monofilament suture. The successfully induced rats were assigned to inosine group (n =32) and model group (n =32) at random. Rats in the inosine group were intraperitoneally administrated with inosine in dose of 100 mg/kg preoperatively, twice a day, 7 days in all. The rats in the control group were injected with the same dose of saline solution by the similar way preoperatively. Each group was randomized into ischemia /reperfusion 2, 6, 12, 24 hours, 2, 3, 7 and 14 days subgroups consisted of 4 rats. The other 4 rats were taken as the sham-operated group, the rats were given the same treatment except for not introduced the filament into the external carotid artery stump, and brain tissue was removed at 2 hours of reperfusion. ② In situ hybridization was performed to examine the expression of cytochrome C mRNA while TUNEL staining was made to characterize apoptosis. ③ The t test was used to compare the difference of measurement data. MAIN OUTCOME MEASURES: ① Neuronal apoptosis in the different regions of the ischemic brain tissue; ② Expression of cytochrome C mRNA in the different regions at different time points after MCAO. RESULTS: All the 68 rats were involved in the analysis of results. ① Neuronal apoptosis: A small number of TUNEL-positive cells were detected in the sham-operated brain and non-ischemic brain. The number of apoptotic cells in the ischemic cortex peaked at 24 hours of reperfusion [(72.00±1.98) cells] and that in the striatum peaked at 2 days [(94.75±3.57) cells], then decreased to the level of sham-operated group at 14 days. Inosine could reduce apoptotic cells from 12 hours to 7 days of reperfusion as compared with the model group (t =6.19-26.67, P < 0.01). ② Cytochrome C mRNA expression: There was weak expression of cytochrome C mRNA in both sham-operated brain and contralateral brain. Cytochrome C was detected at 2 hours of reperfusion in ischemic brain [(25.75±3.50), (39.75±2.49) cells], and strongly increased to a peak at 12 hours and 24 hours of reperfusion in cortex and striatum [(122.50±6.69), (119.25±5.12) cells], respectively. Furthermore, inosine could significantly decrease cytochrome C expression in cortex at 12 hours to 14 days of reperfusion after ischemic reperfusion and that in striatum at 12 hours to 3 days (t =8.67-43.26, P < 0.01). CONCLUSION: Inosine can exert a neuroprotective effect by inhibiting apoptosis and cytochrome C mRNA expression.展开更多
Objective To testify the lung injury induced by cardiopulmonary bypass(CPB) in canine model and observe the influence of CPB on the aquaporin 1 (AQP1) mRNA expression in canine lung. Methods 8 mongrel dogs were used t...Objective To testify the lung injury induced by cardiopulmonary bypass(CPB) in canine model and observe the influence of CPB on the aquaporin 1 (AQP1) mRNA expression in canine lung. Methods 8 mongrel dogs were used to perform the cardiopulmonary bypass. The hearts arrested for 90 minutes with mild hypothermia and rebeated for 6 hours. The hemodynamics,the ratio of lung dry weight and wet weight,the plasmic展开更多
To explore the influence of aprotinin on the mRNA expression of P-selectin and ICAM-1 in lung tissue after ischemic reperfusion.Methods Thirty New Zealand white rabbits were randomly divided into 3 groups:control grou...To explore the influence of aprotinin on the mRNA expression of P-selectin and ICAM-1 in lung tissue after ischemic reperfusion.Methods Thirty New Zealand white rabbits were randomly divided into 3 groups:control group,LPD group and aprotinin group.In situ rabbit lung preservation model was established.In control group,the left lower lung lobe was stored at 10℃ in a specially made lung preservation container for 2 hours and reperfused for another 2 hours.In LPD group and aprotinin group,the left lower lobe was perfused with LPD solution or aprotinin containing LPD solution,respectively,after left lung hilus was clamped.The other procedures were the same as those in control group.The lung tissue was harvested at different time intervals including preclamping lung hilus,2 hours after clamping and 2 hours after reperfusion.The mRNA expression of ICAM-1 and P-selectin in the lung tissue was detected with RT-PCR technique.Results The contents of mRNA of P-selectin at 2 hours after reperfusion in control group and LPD group were significant higher than pre-ischemia and 2 hours after champing the left lung hilus.There was no such significant difference in aprotinin group.The mRNA expression of P-selectin in aprotinin group at 2 hours after reperfusion was significantly lower than that in control group and LPD group.The ICAM-1 mRNA expression at 2 hours after Ischemia and 2 hours after reperfusion in control group and LPD group was significantly higher than the pre-ischemia and its was significantly higher than that in aprotinin group.Conclusion Aprotinin can inhibit the upregulation of the mRNA expression of P selectin and ICAM-1 after ischemia reperfusion in the lung tissue,so the addition of aprotinin in LPD solution may reduce the ischemia reperfusion injury in lung tissue.5 refs,1 tab.展开更多
Extensive use of insecticides on cotton has prompted resistance development in the cotton aphid, Aphis gossypii （Glover） in China, A deltamethrin-selected population of cotton aphids from Xinjiang Uygur Autonomous R...Extensive use of insecticides on cotton has prompted resistance development in the cotton aphid, Aphis gossypii （Glover） in China, A deltamethrin-selected population of cotton aphids from Xinjiang Uygur Autonomous Region, China with 228,59-fold higher resistance to deltamethrin was used to examine how carboxylesterase conferred resistance to this pyrethroid insecticide. The carboxylesterase activity in the deltamethrin-resistant strain was 3.67-, 2,02- and 1.16-fold of the susceptible strain when using α-naphthyl acetate （α-NA）, β-naphthyl acetate （β-NA） and α-naphthyl butyrate （α-NB） as substrates, respectively, Carboxylesterase cDNA was cloned and sequenced from both deltamethrinresistant and susceptible strains. The cDNA contained 1581 bp open reading frames （ORFs） coding a 526 amino acid protein. Only one amino acid substitution （Val^87-Ala） was observed between deltamethrin-resistant and susceptible strains but it is not genetically linked to resistance by the catalytic triad and signature motif analysis. The real-time polymerase chain reaction analysis indicated that the resistant strain had a 6.61-fold higher level of carboxylesterase mRNA than the susceptible strain. The results revealed that up-regulation of the carboxylesterase gene, not modified gene structure, may be responsible for the development of resistance in cotton aphids to deltamethrin.展开更多
BACKGROUND: Hypersecretion of biliary cholesterol is believed to be one of the important causes of lithogenic bile. Sterol carrier protein-2 ( SCP2 ) participates in choles- terol trafficking and metabolism and may pl...BACKGROUND: Hypersecretion of biliary cholesterol is believed to be one of the important causes of lithogenic bile. Sterol carrier protein-2 ( SCP2 ) participates in choles- terol trafficking and metabolism and may play a key role in cholesterol gallstone formation. This study was undertaken to investigate the expression of liver SCP2 mRNA in pa- tients with cholesterol gallstone and those patients with non-cholesterol gallstone. METHODS: The expression of liver SCP2 mRNA was studi- ed in 36 patients with cholesterol gallstone and 30 patients with non-cholesterol gallstone by reverse transcription-poly- merase chain reaction (RT-PCR). RESULT: The expression of SCP2 mRNA was increased more significantly in patients with cholesterol gallstone than in patients with non-cholesterol gallstone. CONCLUSION: The SCP2 gene was overexpressed in pa- tients with cholesterol gallstone, indicating that SCP2 may be one of the important causes of cholesterol gallstone.展开更多
Purpose： Host response to polytrauma occasionally has unpredictable outcomes. Immune response is a major factor influencing patient＇s outcome. This study evaluated the interaction of two main cytokines in immune res...Purpose： Host response to polytrauma occasionally has unpredictable outcomes. Immune response is a major factor influencing patient＇s outcome. This study evaluated the interaction of two main cytokines in immune response after major trauma, specifically interleukin-6 （IL-6） and interleukin-10 （IL-10）. Plasma level of these cytokines is determined by mRNA expression of these cytokines genes which may decide the outcome of polytrauma patients. Methods： This prospective multicenter trial held at four trauma centers enrolled 54 polytrauma patients [Injury Severity Score （ISS） ≥ 16]. Plasma levels and mRNA expression of IL-6 and IL-10 were measured for 5 days after trauma. Clinical evaluation was conducted to observe whether patients endured multiple organ dysfunction syndrome （MODS） and death. MODS evaluation was performed using sequential organ failure assessment （SOFA）. Trauma load which in this study is represented with ISS, plasma level, expression of cytokine genes and patient＇s outcome were examined with correlation test and statistical analysis. Results： The elevated IL-6/IL-10 ratio indicated increased activity of systemic inflammation response, especially pro-inflammation response which bears higher probability of progressing to MODS and death. The decline of IL-6/IL-IO ratio with heavy trauma load （1SS 〉 30） showed that compensatory anti- inflammation response syndrome （CARS） state was more dominant than systemic inflammatory response syndrome （SIRS）, indicating that malfunction and failure of immune system eventually lead to MODS and deaths. The statistical significance in plasma level of cytokines was found in the outcome group which was defined as bearing a low trauma load but mortality. Conclusion： The pattern of cytokine levels in inflammation response has great impact on the outcome of polytrauma patients. Further study at the genetic level is needed to investigate inflammation process which may influence patient＇s outcome.展开更多
基金Supported by JSPS Ronpaku (Dissertation PhD) program (No.NRCT 10726) award by Japan Society for the Promotion of Scince and in collaboration with Kobe University School of Medicine,Kobe,JapanJSPS Asian CORE Program 2012,Nippon Medical Schoolthe Faculty of Medicine,Chiang Mai University,Chiang Mai,Thailand (in part)
文摘AIM: To study gastric mucosal interleukine-8 (IL-8) mRNA expression, the cytotoxin-associated gene A (cagA) mutation, and serum pepsinogen (PG)?I/II ratio related risk in Thai gastric cancer.METHODS: There were consent 134 Thai non-cancer volunteers who underwent endoscopic narrow band imaging examination, and 86 Thais advance gastric cancer patients who underwent endoscopic mucosal biopsies and gastric surgery. Tissue samples were taken by endoscopy with 3 points biopsies. The serum PG?I, II, and Helicobacter pylori (H. pylori) immunoglobulin G (IgG) antibody for H. pylori were tested by enzyme-linked immunosorbent assay technique. The histopathology description of gastric cancer and non-cancer with H. pylori detection was defined with modified Sydney Score System. Gastric mucosal tissue H. pylori DNA was extracted and genotyped for cagA mutation. Tissue IL-8 and cyclooxygenase-2 (COX-2) mRNA expression were conducted by real time relative quantitation polymerase chain reaction. From 17 Japanese advance gastric cancer and 12 benign gastric tissue samples, all were tested for genetic expression with same methods as well as Thai gastric mucosal tissue samples. The multivariate analysis was used for the risk study. Correlation and standardized t-test were done for quantitative data, P value < 0.05 was considered as a statistically significant.RESULTS: There is a high non cagA gene of 86.8 per cent in Thai gastric cancer although there are high yields of the East Asian type in the positive cagA. The H. pylori infection prevalence in this study is reported by combined histopathology and H. pylori IgG antibody test with 77.1% and 97.4% of sensitivity and specificity, respectively. The serum PG?I/II ratio in gastric cancer is significantly lower than in the non-cancer group, P = 0.045. The serum PG?I/II ratio of less than 3.0 and IL-8 mRNA expression ≥ 100 or log10 ≥ 2 are significant cut off risk differences between Thai cancer and non-cancer, P = 0.03 and P < 0.001, respectively. There is a significantly lower PGI/II ratio in Japanese than that in Thai gastric cancer, P = 0.026. Serum PG?I/II ratio at cut off less than 3.0 and IL-8 mRNA expression Raw RQ > 100 or log10 > 2 are significantly difference between Thai cancer group when compared to non-cancer group, P = 0.013 and P < 0.001, respectively. In the correlation study, low PG?I/II ratio does not associate with chronic atrophic gastritis severity score in Thais non-cancer cases. However, there is a trend, but not significant convert correlation between IL-8 mRNA expression level and low PG?I/II ratio in Thai positive H. pylori infection. The high expression of IL-8 gene demonstrates a poorer prognosis by stage and histology.CONCLUSION:Predominant gastric mucosal IL-8 mRNA expression level, H. pylori infection, and low PG?I/II ratio are relative risks for Thai gastric cancer without correlation with cagA mutation.
基金Supported by the Molecular Biology Program (Grant No.21407)Laboratory Medicine Center-LMC, University Hospital Linkoping, Swedenthe Development Foundation of Region Skane, Sweden
文摘AIM： To investigate the expression of gastrin-releasing peptide （GRP） and GRP-receptor mRNA in non-tumor tissues of the human esophagus, gastrointestinal tract, pancreas and gallbladder using molecular biology techniques. METHODS： Poly A^＋ mRNA was isolated from total RNA extracts using an automated nucleic acid extractor and, subsequently, converted into single-stranded cDNA （sscDNA）. PCR amplifications were carried out using genespecific GRP and GRP-receptor primers. The specificity of the PCR amplicons was further confirmed by Southern blot analyses using gene-specific GRP and GRP-receptor hybridization probes. RESULTS： Expression of GRP and GRP-receptor mRNA was detected at various levels in nearly all segments of the non-tumor specimens analysed, except the gallbladder. In most of the biopsy specimens, coexpression of both GRP and GRP-receptor mRNA appeared to take place. However, expression of GRP mRNA was more prominent than was GRP-receptor mRNA. CONCLUSION： GRP and GRP-receptor mRNAs are expressed throughout the gastrointestinal tract and provides information for the future mapping and determination of its physiological importance in normal and tumor cells.
基金The National Natural Science Foundation of China under contract No. 40976096the National Department Public Benefit Research Foundation of China under contract No. 200903029the National High Technology Research and Development Program of China (863 Program) under contract No. 2006AA10A405
文摘Mammalian macrophage migration inhibitory factor （MIF） plays an important role as an indispensable mediator in the pathogenesis of inflammatory disease like septicemia, but little is known about the role of MIF homologue in fish septicemia. The authors have cloned the MIF homologue in large yellow croaker Pseudosciaena crocea （LycMIF） using RACE approach. The full-length cDNA of LycMIF was 634 bases and contained an ORF of 345 bases encoding a protein of 115 amino acid residues. As demonstrated by RT-PCR and QRT-PCR assay, MIF mRNAs were constitutively expressed in 11 selected tissues and were abundant in brain and liver. Moreover, the LycMIF transcripts in the liver and head kidney were responsive to bacteria infection and could be significantly up-regulated. Our results provide the first direct evidence that fish MIF was implicated in pathogenesis of fish vibrosis and play an important role in response to bacteria infection.
基金supported by the National Natural Science Foundation of China(31301967)the Natural Science Foundation of Jiangsu Province,China(BK20161322)+4 种基金the projects of Key Laboratory for Poultry Genetics and Breeding of Jiangsu Province(JQLAB-ZZ-201703)the Major Breeding Programs in Jiangsu Province,China(PZCZ201728)the earmarked fund for China Agriculture Research System(CARS-41)the Independent Scientific Foundation of Public Welfare Scientific Institutes in Jiangsu Province,China(BM2018026)the Open Projects of Key Laboratory of Chicken Genetics and Breeding,Ministry of Agriculture and Rural Affairs of China(CGB-201704)。
文摘Previous studies on mammals showed that peroxisome proliferator-activated receptor gamma coactivator-1α(PGC-1α)played a prominent role in regulating muscle fiber type transition and composition.However,the role of PGC-1αin chicken muscle has seldom been explored.To investigate the effect of PGC-1αon chicken skeletal muscles in this study,the PGC-1αgene was overexpressed or silenced in chicken primary myoblasts by using lentivirus,and then the effects of the PGC-1αgene overexpression and knockdown on the mRNA expression profile of genes related to myofiber type specificity were examined during fiber formation.The results showed that overexpression of PGC-1αfrom proliferation to differentiation was accompanied by the up-regulated expression of Pax7,MyoD,and CnAα,which was significantly(P<0.01)increased after one day of transfection(1 I).The enhancement of MyoG,MEF2 c,and MyHC SM expression lagged,which was improved significantly(P<0.01)after four days of transfection(1 I3 D).Overexpression of PGC-1αdecreased(P<0.01)the MyHC FWM expression after four days of transfection(1 I3 D),and it had no significant impact(P>0.05)on the expression of CnB1,NFATc3,and MyHC FRM during myofiber formation.The effective silence(P<0.01)of PGC-1αby lentivirus mediating short hairpin RNA(shRNA)was detected after four days of transfection(1 I3 D)in cultures,and the lack of its function in chicken primary myoblasts significantly(P<0.01)down-regulated the expression of Pax7,MyoD,CnAα,MyoG,MEF2 c,and MyHC SM,significantly(P<0.01)up-regulated the expression of MyHC FWM,and had no significant impact(P>0.05)on the expression of CnB1,NFATc3,and MyHC FRM.These results indicated that the role of PGC-1αin regulating the fiber type specificity of chicken skeletal muscles might be similar to that in mammals,which interplayed with key genes related to myocyte differentiation and calcineurin signaling pathway.
基金supported by a grant from the National Natural Science Foundation of China (30500360)the Open Topic of State Key Laboratory of Agricultural Biotechnology (2009SKLAB07-2)
文摘The H19 gene, which is imprinted with preferential expression from the maternal allele, was one of the first identified imprinting genes in mammals. Recent studies revealed that correct imprinting of the H19 gene plays a vital role in human spermatogenesis. To investigate whether imprinting defects were associated with the hybrid sterility of male cattle-yak, the methylation patterns of the H19 imprinting control region （ICR） and H19 mRNA expression in the testes of cattle-yak, yak, and cattle were examined. The results showed that the 3rd CCCTC-binding factor （CTCF） site of the H19 ICR was significantly hypomethylated in the testes of cattle-yak compared with yak or cattle. As expected, H19 was expressed at a significantly higher level in cattle-yak than in yak or cattle. These results suggest that imprinting defects of the CTCF- binding site in the HI9 ICR were possibly associated with disturbed spermatogenesis of male cattle-yak. Thus, we propose that disorders in H19 imprinting, resulting in an increased H19 mRNA expression, might contribute to the sterility of F1 male hybrids between cattle and yak.
基金supported by the“863"Prijetof China under contract Nos 2001AA628180 and 2002AA626020.
文摘A homologue of the lower vertebrates translationally controlled tumor protein (TCTP) was cloned from the marine fish Japanese sea perch (Lateolabrax japonicus) by the technology of homology cloning. The full-length cDNA sequence of the sea perch TCTP gene contained a 5' untranslated region (UTR) of 47 bp, a 3' UTR of 433 bp, and a putative open reading frame (ORF) of 510 bp encoding a polypeptide of 170 amino acids. The deduced amino acid sequence of the sea perch TCTP gene showed a high similarity to that of zebrafish, rohu, rabbit, chicken and human. Sequence analysis revealed there were a signature sequence of TCTP family, an N-glycosylation site, and five Casein kinase phosphorylation sites in the sea perch TCTP. The temporal expression of TCTP genes in healthy and lipopolysaccharide (LPS) challenged fishes was measured by semi-quantitative reverse transcription-PCR (RT-PCR). The results indicated that LPS could up-regulate the expression of sea perch TCTP in the examined tissues, including head-kidney, spleen and liver.
文摘Background:Maternal nutrition during gestation affects fetal development,which has long-term programming effects on offspring postnatal growth performance.With a critical role in protein and lipid metabolism,essential fatty acids can influence the development of muscle and adipose tissue.The experiment investigated the effects of late gestation supplements(77 d prepartum),either rich in saturated and monounsaturated fatty acids(CON;155 g/cow/d EnerGII)or polyunsaturated fatty acids(PUFA;80 g/cow/d Strata and 80 g/cow/d Prequel),on cow performance and subsequent calf growth performance as well as mRNA expression in longissimus muscle(LM)and subcutaneous adipose tissue at birth and weaning.Results:There was no difference(P≥0.34)in cow body weight(BW)or body condition score from presupplementation through weaning.Relative concentrations of C18:3n-3 and C20:4n-6 decreased(P≤0.05)to a greater extent from mid-supplementation to calving for PUFA compared with CON cows.Cow plasma C20:0,C20:5n-3,and C22:6n-3 were increased(P≤0.01)in PUFA during supplementation period.At birth,PUFA steers had greater(P=0.01)plasma C20:5n-3.No differences(P≥0.33)were detected in steer birth BW or dam milk production,however,CON steers tended(P=0.06)to have greater pre-weaning average daily gain and had greater(P=0.05)weaning BW compared with PUFA.For mRNA expression in steers:MYH7 and C/EBPβin LM increased(P≤0.04)to a greater extent from birth to weaning for PUFA compared with CON;MYF5 in LM and C/EBPβin adipose tissue tended(P≤0.08)to decrease more from birth to weaning for CON compared with PUFA;SCD in PUFA adipose tissue tended(P=0.08)to decrease to a greater extent from birth to weaning than CON.In addition,maternal PUFA supplementation tended(P=0.08)to decrease MYOG mRNA expression in LM and decreased(P=0.02)ZFP423 in adipose tissue during the pre-weaning stage.Conclusions:Late gestation PUFA supplementation decreased pre-weaning growth performance of the subsequent steer progeny compared with CON supplementation,which could have been a result of downregulated mRNA expression of myogenic genes during pre-weaning period.
基金supported by the Agriculture Profession Item (3-52)by the earmarked fund for Modern Agro-Industry Technology Research System of the Ministry of Agriculture of the P.R.China (nycytx-44)
文摘The effects of dietary lysine on production performance,serum concentrations of metabolites,growth hormone （GH）,insulin-like growth factor-I （IGF-I） and IGF-I mRNA expression in growing rabbits were examined.One hundred weaned New Zealand rabbits were allocated to individual cages and randomly offered a diet containing 5.5 （L1）,6.5 （L2）,7.5 （L3）,8.5 （L4）,or 9.5 g （L5） lysine per kg diet.The results showed that the average daily gain （ADG） of the rabbits from L3,L4 or L5 was higher than those from L1 or L2 （P 〈 0.05）.The feed gain ratio （F/G） in the rabbits from L4 or L5 was lower than those from L1 or L2 （P 〈 0.05）.Dietary lysine did not affect serum concentrations of total protein （TP）,glucose,insulin （INS）,and growth hormone （GH） （P 〉 0.05）.The quadratic effects of lysine on the serum urea nitrogen （SUN） concentration was detected （P = 0.035）.Serum IGF-I concentrations had a trend to increase quadratically with the increasing dietary lysine （P = 0.07）.A significant correlation was found between serum IGF-I concentrations （x,ng mL-1） and ADG （y,g kg-1）： y = -0.017x2 ＋ 1.984x ＋ 20.87 （R2 = 0.8982,P = 0.003）.The relative abundance of hepatic and muscular IGF-I mRNA tended to increase with increasing dietary lysine levels （P = 0.053 and 0.082,respectively）.Providing the diets mainly consisted of corn,wheat bran and peanut vine,the most appropriate dietary lysine level for growing meat rabbits from weaning to 70 d old was found to be 8.5 g kg-1,and IGF-I may be an important factor controlling growth of weaned rabbits.
基金Supported by a research grant from the National Science Council, Taipei, Taiwan, China. No. NSC 92-2320-B-038-029
文摘AIM： To investigate the effects of arginine （Arg）-enriched diets before sepsis and/or Arg-containing total parenteral nutrition （TPN） after sepsis or both on cytokine mRNA expression levels in splenocytes of rats with gut-derived sepsis. METHODS： Rats were assigned to four experimental groups. Groups 1 and 2 were fed with a semipurified diet, while groups 3 and 4 had part of the casein replaced by Arg which provided 2% of the total calories. After the rats were fed with these diets for 10 d, sepsis was induced by cecal ligation and puncture （CLP）, at the same time an internal jugular vein was cannulated. All rats were maintained on TPN for 3 d. Groups 1 and 3 were infused with conventional TPN, while groups 2 and 4 were supplemented with Arg which provided 2% of the total calories in the TPN solution. All rats were killed 3 d after CLP to examine their splenocyte subpopulation distribution and cytokine expression levels. RESULTS： Plasma interleukin （IL）-2, IL-4, tumor necrosis factor-α （TNF-α） and interferon （IFN-γ） were not detectable 3 d after CL.P. There were no differences in the distributions of CD45Ra＋, CD3＋, CD4＋, and CD8＋ cells in whole blood and splenocytes among the four groups. The splenocyte IL-2 mRNA expression in the Arg-supplemented groups was significantly higher than that in group 1. IL-4 mRNA expression in groups 3 and 4 was significantly higher than that in groups 1 and 2. The mRNA expression of IL-10 and IFN-γ was significantly higher in group 4 than in the other three groups. There was no difference in TNF-α mRNA expression among thefour groups CONCLUSION： The influence of Arg on the whole blood and splenic lymphocyte subpopulation distribution is not obvious. However, Arg administration, especially before and after CLP, significantly enhances the mRNA expression levels of Thl and Th2 cytokines in the spleen of rats with gut-derived sepsis.
基金supported by grants from the Scientific Research Foundation for Returned Scholars of the Ministry of Education of ChinaPre-research Foundation of Wuhan University,China(No.301270050)
文摘Summary： In adult liver, CYP3A4 plays an important role in the metabolism of a wide range of en- dogenous and exogenous compounds. To investigate whether there is a single nucleotide polymorphism （SNP） of CYP3A4 intron 2 in the liver and its effects on the mRNA expression and enzymatic activity of CYP3A4, genomic DNA was extracted from 96 liver tissue samples obtained from patients who had undergone liver surgery. An SNP of CYP3A4 intron 2 was identified by polymerase chain reaction （PCR）-single-strand confirmation polymorphism and DNA sequencing. The mRNA expression of CYP3A4 was determined by the fluorescence quantitative PCR technique. The enzymatic activity of CYP3A4 was measured using erythromycin and testosterone as probe substrates. Twelve patients were found to have the SNP/T4127G CYP3A4 within intron 2. The mRNA levels of CYP3A4 in wild-type and SNP/T4127G samples were 2.62±1.09 and 2.79±1.63, respectively （P〉0.05）. Erythromycin N-demethylase activity in wild-type and SNP/T4127G samples were 121.2±32.8 and 124.7±61.6 nmol·mg^-1min^-1, respectively （P〉0.05）. The activity of testosterone 613-hydroxylase was significantly different between wild-type （648±173 pmol·mg^-1·min^-1） and SNP/T4127G samples （540-4-196 pmol.mg-l-minl; P〈0.05）. In conclusion, the SNP/T4127G of CYP3A4 intron 2 exists in the liver. This SNP does not affect the mRNA expression of CYP3A4 but significantly decreases the hepatic micro- somal testosterone 613-hydroxylase activity of CYP3A4. Furthermore, this study indicates that the ap- propriate selection of probe substrates is very important in studying the relationship between the geno- type and phenotype of CYP3A4.
基金Supported by the National Natural Science Foundation of China (No.41206144)the National High Technology Research and Development Program of China (863 Program) (No. 2008AA100805)
文摘AMP deaminase catalyzes the conversion of AMP into IMP and ammonia. In the present study, a full-length cDNA of AMPD1 from skeletal muscle of Japanese flounder Paralichthys olivaceus was cloned and characterized. The 2 526 bp cDNA contains a 5'-UTR of 78 bp, a 3'-UTR of 237 bp and an open reading frame (ORF) of 2 211 bp, which encodes a protein of 736 amino acids. The predicted protein contains a highly conserved AMP deaminase motif (SLSTDDP) and an ATP-binding site sequence (EPLMEEYAIAAQVFK). Phylogenetic analysis showed that the AMPD1 and AMPD3 genes originate from the same branch, but are evolutionarily distant from the AMPD2 gene. RT-PCR showed that the flounder AMPD1 gene was expressed only in skeletal muscle. QRT-PCR analysis revealed a statistically significant 2.54 fold higher level of AMPD1 mRNA in adult muscle (750±40 g) compared with juvenile muscle (7.5±2 g) (P<0.05). HPLC analysis showed that the IMP content in adult muscle (3.35±0.21 mg/g) was also statistically significantly higher than in juvenile muscle (1.08±0.04 mg/g) (P<0.05). There is a direct relationship between the AMPD1 gene expression level and IMP content in the skeletal muscle of juvenile and adult flounders. These results may provide useful information for quality improvement and molecular breeding of aquatic animals.
基金the National Natural Science Foundation of China, No. 30171188
文摘BACKGROUND： Somatostatin is abundant in the hypothalamus, cerebral cortex, limbic system, and mesencephalon. Somatostatin mRNA expression in the brain of rats with spleen deficiency is noticeably reduced, as well as attenuation of cognitive function. OBJECTIVE： To observe the interventional effect of Guipi decoction on somatostatin level and somatostatin receptor 1 （SSTRl） mRNA expression in different encephalic regions of rats with spleen deficiency, and to compare the interventional effects of Guipi decoction, Chaihu Shugan powder, and Tianwang Buxin pellet. DESIGN： A randomized controlled observation. SETTING： Basic Medical College, Beijing University of Traditional Chinese Medicine. MATERIALS： Fifty adult Wistar male rats, of clean grade, weighing （160 ± 10） g, were provided by Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd. The protocol was performed in accordance with ethical guidelines for the use and care of animals. Somatostatin 1 polyclonal anti-rabbit antibody and SSTRl in situ hybridization kit were provided by Department of Neuroanatomy, Shanghai Second Military Medical University of Chinese PLA. The drug for developing rat models of spleen deficiency was composed of Dahuang, Houpu and Zhishi, and prepared at 2：1：1. Guipi decoction, Chaihu Shugan powder, and Tianwang Buxin pellet recipes were made according to previous studies. METHODS： This study was performed at the Basic Medical College, Beijing University of Traditional Chinese Medicine from March 2002 to March 2005. The rats were randomly divided into 5 groups, with 10 rats in each group： normal, model, Guipi decoction, Chaihu Shugan powd.er, and Tianwang Buxin pellet groups. Rat models of the latter 4 groups were developed by methods of purgation with bitter and cold nature drugs, improper diet, and overstrain. The rats received 7.5 g/kg of the drugs each morning and were fasted every other day, but were allowed free access to water at all times. The rats were forced to swim in 25 ℃ water until fatigued. Rats in the normal group were intragastrically administered the same amount of normal saline. Rats in the Guipi decoction, Chaihu Shugan powder, and Tianwang Buxin pellet groups were intragastrically administered 7.5 g/kg Guipi decoction, Chaihu Shugan powder, and Tianwang Buxin pellet, respectively, every afternoon. All rats were treated for 6 weeks. MAIN OUTCOME MEASURES： Somatostatin protein and SSTRI mRNA expression in the ventral nucleus of hypothalamus, hippocampal CAl region, and cortex of prefrontal lobe were determined by immunohistochemistry and in situ hybridization, respectively. RESULTS： Fifty rats were included in the final analysis. In the model group, expression of somatostatin protein and SSTRl mRNA in the ventral nucleus of hypothalamus, hippocampal CAl region, and cortex of prefrontal lobe were significantly less than in the normal group （P 〈 0.01）. Above-mentioned indices were identical in the Chaihu Shugan powder and model groups. However, expression of somatostatin protein and SSTRl mRNA were significantly higher in the Guipi decoction group compared to model group （P 〈 0.01）. In the Tianwang Buxin pellet group, SSTRl mRNA expression in rat ventral nucleus of hypothalamus and somatostatin level in rat hippocampal CAl region and cortex of prefrontal lobe, as well as ventral nucleus of hypothalamus, were significantly higher compared to model group （P 〈 0.01 ）. CONCLUSION： Somatostatin level and SSTRl mRNA expression in rats with spleen deficiency were lower than in normal rats. Guipi decoction and Tianwang Buxin pellet up-regulated somatostatin level and SSTRl mRNA expression.
文摘Objective: To look for additional markers of molecular biology response to anastrozole, a new aromatase inhibitor, on the growth and mRNA expression level of MCF-7 cell. Methods: We investigated the effect of anastrzole on growth and gene expression in the human breast cancer cell line MCF-7 and compared with the most widely used antiestrogen tamoxifen. We chose 4 genes to examine regulation of gene expression of estrogen regulated genes: PR A, PR B, ErbB-2 and cyclin D1. Results: Compared with the tamoxifen, a statistically significant growth inhibition was observed with anastrozole. The PR A, PR B and cyclin D1 mRNA level in anastrozole treated cells was sigificantly below the level in tamoxifen treated cells (P<0. 05). They had agonistic effect on ErbB gene (P>0. 05). Conclusion: The third generation of aromatase inhibitors anastrozole exert more inhibit function in some expression of estrogen regulated genes than tomoxifen in MCF-7 cell line.
基金the Natural Science Fund of Shandong Province, No.Y2001C04
文摘BACKGROUND: It has been demonstrated that adenosine can induce glial cell to release cytochrome C, enhance expression of apoptotic gene bax, inhibit anti-apoptotic gene bcl-2, and activate caspase-3 to apoptosis; Whereas inosine can inhibit neuronal apoptosis which is similar to bcl-2. OBJECTIVE: To observe the effects of inosine on neuronal apoptosis and expression of cytochrome C mRNA in rats after focal cerebral ischemia/reperfusion, and analyze the pathway of its neuroprotective effect. DESIGN: A randomised controlled animal trial. SETTINGS: Department of Neurology, Rongcheng Second People's Hospital; Department of Neurology, Affiliated Union Hospital, Tongji Medical College, Huazhong University of Science and Technology. MATERIALS: Sixty-eight rats, weighing 230-280 g and clean grade, were used. TdT-mediated dUTP-biotin nick end labeling (TUNEL) and cytochrome C mRNA in situ hybridization kits and DAB staining kit were purchased from Wuhan Boster Biological Co., Ltd.; Inosine injection [200 mg (2 mL) each] from Qingdao First Pharmaceutical Factory. METHODS: The experiment was accomplished in the animal experimental center in Tongji Medical College of Huazhong University of Science and Technology from December 2003 to June 2005. ① Sixty-four rats were made into focal ischemia by middle cerebral artery occlusion (MCAO) with a nylon monofilament suture. The successfully induced rats were assigned to inosine group (n =32) and model group (n =32) at random. Rats in the inosine group were intraperitoneally administrated with inosine in dose of 100 mg/kg preoperatively, twice a day, 7 days in all. The rats in the control group were injected with the same dose of saline solution by the similar way preoperatively. Each group was randomized into ischemia /reperfusion 2, 6, 12, 24 hours, 2, 3, 7 and 14 days subgroups consisted of 4 rats. The other 4 rats were taken as the sham-operated group, the rats were given the same treatment except for not introduced the filament into the external carotid artery stump, and brain tissue was removed at 2 hours of reperfusion. ② In situ hybridization was performed to examine the expression of cytochrome C mRNA while TUNEL staining was made to characterize apoptosis. ③ The t test was used to compare the difference of measurement data. MAIN OUTCOME MEASURES: ① Neuronal apoptosis in the different regions of the ischemic brain tissue; ② Expression of cytochrome C mRNA in the different regions at different time points after MCAO. RESULTS: All the 68 rats were involved in the analysis of results. ① Neuronal apoptosis: A small number of TUNEL-positive cells were detected in the sham-operated brain and non-ischemic brain. The number of apoptotic cells in the ischemic cortex peaked at 24 hours of reperfusion [(72.00±1.98) cells] and that in the striatum peaked at 2 days [(94.75±3.57) cells], then decreased to the level of sham-operated group at 14 days. Inosine could reduce apoptotic cells from 12 hours to 7 days of reperfusion as compared with the model group (t =6.19-26.67, P < 0.01). ② Cytochrome C mRNA expression: There was weak expression of cytochrome C mRNA in both sham-operated brain and contralateral brain. Cytochrome C was detected at 2 hours of reperfusion in ischemic brain [(25.75±3.50), (39.75±2.49) cells], and strongly increased to a peak at 12 hours and 24 hours of reperfusion in cortex and striatum [(122.50±6.69), (119.25±5.12) cells], respectively. Furthermore, inosine could significantly decrease cytochrome C expression in cortex at 12 hours to 14 days of reperfusion after ischemic reperfusion and that in striatum at 12 hours to 3 days (t =8.67-43.26, P < 0.01). CONCLUSION: Inosine can exert a neuroprotective effect by inhibiting apoptosis and cytochrome C mRNA expression.
文摘Objective To testify the lung injury induced by cardiopulmonary bypass(CPB) in canine model and observe the influence of CPB on the aquaporin 1 (AQP1) mRNA expression in canine lung. Methods 8 mongrel dogs were used to perform the cardiopulmonary bypass. The hearts arrested for 90 minutes with mild hypothermia and rebeated for 6 hours. The hemodynamics,the ratio of lung dry weight and wet weight,the plasmic
文摘To explore the influence of aprotinin on the mRNA expression of P-selectin and ICAM-1 in lung tissue after ischemic reperfusion.Methods Thirty New Zealand white rabbits were randomly divided into 3 groups:control group,LPD group and aprotinin group.In situ rabbit lung preservation model was established.In control group,the left lower lung lobe was stored at 10℃ in a specially made lung preservation container for 2 hours and reperfused for another 2 hours.In LPD group and aprotinin group,the left lower lobe was perfused with LPD solution or aprotinin containing LPD solution,respectively,after left lung hilus was clamped.The other procedures were the same as those in control group.The lung tissue was harvested at different time intervals including preclamping lung hilus,2 hours after clamping and 2 hours after reperfusion.The mRNA expression of ICAM-1 and P-selectin in the lung tissue was detected with RT-PCR technique.Results The contents of mRNA of P-selectin at 2 hours after reperfusion in control group and LPD group were significant higher than pre-ischemia and 2 hours after champing the left lung hilus.There was no such significant difference in aprotinin group.The mRNA expression of P-selectin in aprotinin group at 2 hours after reperfusion was significantly lower than that in control group and LPD group.The ICAM-1 mRNA expression at 2 hours after Ischemia and 2 hours after reperfusion in control group and LPD group was significantly higher than the pre-ischemia and its was significantly higher than that in aprotinin group.Conclusion Aprotinin can inhibit the upregulation of the mRNA expression of P selectin and ICAM-1 after ischemia reperfusion in the lung tissue,so the addition of aprotinin in LPD solution may reduce the ischemia reperfusion injury in lung tissue.5 refs,1 tab.
文摘Extensive use of insecticides on cotton has prompted resistance development in the cotton aphid, Aphis gossypii （Glover） in China, A deltamethrin-selected population of cotton aphids from Xinjiang Uygur Autonomous Region, China with 228,59-fold higher resistance to deltamethrin was used to examine how carboxylesterase conferred resistance to this pyrethroid insecticide. The carboxylesterase activity in the deltamethrin-resistant strain was 3.67-, 2,02- and 1.16-fold of the susceptible strain when using α-naphthyl acetate （α-NA）, β-naphthyl acetate （β-NA） and α-naphthyl butyrate （α-NB） as substrates, respectively, Carboxylesterase cDNA was cloned and sequenced from both deltamethrinresistant and susceptible strains. The cDNA contained 1581 bp open reading frames （ORFs） coding a 526 amino acid protein. Only one amino acid substitution （Val^87-Ala） was observed between deltamethrin-resistant and susceptible strains but it is not genetically linked to resistance by the catalytic triad and signature motif analysis. The real-time polymerase chain reaction analysis indicated that the resistant strain had a 6.61-fold higher level of carboxylesterase mRNA than the susceptible strain. The results revealed that up-regulation of the carboxylesterase gene, not modified gene structure, may be responsible for the development of resistance in cotton aphids to deltamethrin.
基金This study was supported by a grant from the Health Bureau of Tianjin, China(No. 00kyzd8).
文摘BACKGROUND: Hypersecretion of biliary cholesterol is believed to be one of the important causes of lithogenic bile. Sterol carrier protein-2 ( SCP2 ) participates in choles- terol trafficking and metabolism and may play a key role in cholesterol gallstone formation. This study was undertaken to investigate the expression of liver SCP2 mRNA in pa- tients with cholesterol gallstone and those patients with non-cholesterol gallstone. METHODS: The expression of liver SCP2 mRNA was studi- ed in 36 patients with cholesterol gallstone and 30 patients with non-cholesterol gallstone by reverse transcription-poly- merase chain reaction (RT-PCR). RESULT: The expression of SCP2 mRNA was increased more significantly in patients with cholesterol gallstone than in patients with non-cholesterol gallstone. CONCLUSION: The SCP2 gene was overexpressed in pa- tients with cholesterol gallstone, indicating that SCP2 may be one of the important causes of cholesterol gallstone.
文摘Purpose： Host response to polytrauma occasionally has unpredictable outcomes. Immune response is a major factor influencing patient＇s outcome. This study evaluated the interaction of two main cytokines in immune response after major trauma, specifically interleukin-6 （IL-6） and interleukin-10 （IL-10）. Plasma level of these cytokines is determined by mRNA expression of these cytokines genes which may decide the outcome of polytrauma patients. Methods： This prospective multicenter trial held at four trauma centers enrolled 54 polytrauma patients [Injury Severity Score （ISS） ≥ 16]. Plasma levels and mRNA expression of IL-6 and IL-10 were measured for 5 days after trauma. Clinical evaluation was conducted to observe whether patients endured multiple organ dysfunction syndrome （MODS） and death. MODS evaluation was performed using sequential organ failure assessment （SOFA）. Trauma load which in this study is represented with ISS, plasma level, expression of cytokine genes and patient＇s outcome were examined with correlation test and statistical analysis. Results： The elevated IL-6/IL-10 ratio indicated increased activity of systemic inflammation response, especially pro-inflammation response which bears higher probability of progressing to MODS and death. The decline of IL-6/IL-IO ratio with heavy trauma load （1SS 〉 30） showed that compensatory anti- inflammation response syndrome （CARS） state was more dominant than systemic inflammatory response syndrome （SIRS）, indicating that malfunction and failure of immune system eventually lead to MODS and deaths. The statistical significance in plasma level of cytokines was found in the outcome group which was defined as bearing a low trauma load but mortality. Conclusion： The pattern of cytokine levels in inflammation response has great impact on the outcome of polytrauma patients. Further study at the genetic level is needed to investigate inflammation process which may influence patient＇s outcome.
