Human growth hormone 1 (GH1) gene expression: complex haplotype-dependent influence of polymorphic variation in the proximal promoter and locus control region

The proximal promoter region of the human pituitary expressed growth hormone (GH1) gene is highly polymorphic, containing at least 15 single nucleotide polymorphisms (SNPs). This variation is manifest in 40 different haplotypes, the high diversity being explicable in terms of gene conversion, recurrent mutation, and selection. Functional analysis showed that 12 haplotypes were associated with a significantly reduced level of reporter gene expression whereas 10 haplotypes were associated with a significantly increased level. The former tend to be more prevalent in the general population than the latter (p<0.01), possibly as a consequence of selection. Although individual SNPs contributed to promoter strength in a highly interactive and non-additive fashion, haplotype partitioning was successful in identifying six SNPs as major determinants of GH1 gene expression. The prediction and functional testing of hitherto unobserved super-maximal and sub-minimal promoter haplotypes was then used to test the efficacy of the haplotype partitioning approach. Electrophoretic mobility shift assays demonstrated that five SNP sites exhibit allele-specific protein binding. An association was noted between adult height and the mean in vitro expression value corresponding to an individual's GH1 promoter haplotype combination (p=0.028) although only 3.3% of the variance of adult height was found to be explicable by reference to this parameter. Three additional SNPs, identified within sites I and II of the upstream locus control region (LCR), were ascribed to three distinct LCR haplotypes. A series of LCR-GH1 proximal promoter constructs were used to demonstrate that 1) the LCR enhanced proximal promoter activity by up to 2.8-fold depending upon proximal promoter haplotype, and that 2) the activity of a given proximal promoter haplotype was also differentially enhanced by different LCR haplotypes. The genetic basis of inter-individual differences in GH1 gene expression thus appears to be extremely complex.     

大学生性行为及性道德观发展研究

目的 了解当代大学生性行为及性道德观念的发展变化。方法 自制问卷，对全国9所高校的2000余名大学生进行调查，并用SPSS10．0对结果进行统计处理。结果 大学生性交行为发生率呈逐年上升趋势，且随年级上升而增加；大学生认可性交行为道德观的5个雏度，但各年级之间有差异；大学生边缘性行为和独自性行为发生率随年级显著上升，认可度也随年级增加而上升；大学生对同性性接触基本持排斥态度，但排斥态度则趋于下降。结论 大学生性行为和性道德随年级上升而明显变化，应加强学校教育和引导。     

Prognostic factors of the long-term survival after transjugular intrahepatic portosystemic shunt in the treatment of gastric and esophageal variceal bleeding

Transjugular intrahepatic portosystemic shunt (TIPSS) is a promising method of treatment for gastric or esophageal variceal bleeding. This study was performed to determine the prognostic factors contributing to the survival of patients after TIPSS for gastric or esophageal variceal bleeding. One hundred and fifty-five patients who underwent TIPSS between September 1991 and March 2001 were followed up by clinical examination, upper gastrointestinal endoscopy, and Duplex sonography. The mean portohepatic pressure gradient prior to TIPSS was 20.5+/-9.93 mmHg and dropped to 10.7+/-6.62 mmHg after TIPSS (p<0.001). The cumulative survival rate was 75.1% at 6 months, 66.6% at 1 yr, 58.4% at 2 yr, and 38.1% at 5 yr. Survival after TIPSS was inversely related to the Child-Pugh classification (p<0.05). The rebleeding rate was 18.3% at 6 months, 21.0% at 1 yr, 32.8% at 2 yr, and 53.1% at 5 yr. The causes of deaths were hepatic failure (53.5%), recurrent variceal bleeding (11.6%), pneumonia (4.6%), sepsis (3.5%), hepatic encephalopathy (2.3%), and unknown (17.4%). Multivariate analysis (Cox proportional hazard model) revealed that the Child-Pugh classification and age were statistically significant independent prognostic factors. In conclusion, TIPSS is an effective method of treatment for variceal bleeding in cases where other treatment modalities including endoscopic therapy are unsuccessful and the most important prognostic factors are preprocedural hepatic reserve (Child-Pugh class) and age.     

非受体型酪氨酸激酶-Syk蛋白的提取与纯化

目的 :从转染syk基因的Sf2 1细胞中提取、纯化免疫相关因子Syk蛋白。方法 :将syk基因转染Sf2 1细胞 ,于 2 8℃培养 48h ,收集细胞 ,用超声波破碎仪裂解细胞 ,提取裂解液中总蛋白 ,用Yellow 3凝胶和Toyopearl AF Heptin 650M凝胶层析柱分离、纯化。层析液中的Syk蛋白存在和性质 ,用SDS PAGE、免疫印迹实验和等电聚焦实验鉴定。结果 :从 2 5亿个Sf2 1细胞裂解液中提取了含有Syk的 2 2 5mg蛋白质。经Yellow 3凝胶层析分离 ,得到两个亚种的Syk蛋白 ,相对分子质量 (Mr)均为72× 10 3 。进一步用Toyopearl AF Heptin 650M凝胶层析纯化后 ,得到两个纯的Syk蛋白 ,SDS PAGE、免疫印迹实验结果显示 ,两种Syk的Mr 均为 72× 10 3,与Syk的理论相对分子质量吻合。但等电聚焦实验显示 ,这两种Syk蛋白成分具有不同的pI值。结论 :从 2 5亿个转染syk基因的Sf2 1细胞中纯化出8mgSyk蛋白 ,纯度高于 95%。这两种Syk的Mr 虽然相同 ,但具有不同的pI值 ,是两个亚种。这些Syk可用于研究Syk的作用机制、抗Syk抗体的制备和Syk诊断试剂盒的制备等     

Safety and efficacy of Salmonella gallinarum 9R vaccine in young laying chickens.

Salmonella enterica subsp. enterica serovar Gallinarum (S. gallinarum) is the agent of fowl typhoid, and the 9R vaccine is a commercially available, live vaccine for the prevention of fowl typhoid. The aim of this study was to assess the safety and efficacy of the 9R vaccine in young chickens. The mean weights of 5-week-old chickens vaccinated with one and 10 doses at 2 weeks old were 450.3+/-33.83 g and 446.8+/-35.68 g, respectively, which were statistically lower (P<0.05) than the mean weight (475.5+/-44.17 g) of the control group. Using the same procedure, the mean weights of chickens vaccinated with one and 10 doses at the age of 4 and 6 weeks were 721.3+/-64.03 g and 723.7+/-63.92 g, and 1114.2+/-92.21 g and 1078.27+/-68.93 g, respectively. Compared with the mean weights (725.7+/-49.50 g and 1104.3+/-92.34 g, respectively) of the control groups, there was no difference in terms of statistical significance (P < 0.05). In addition, all vaccinated birds showed no clinical signs and survived the time course of the experiment. When all chickens were challenged with the wild-type S. gallinarum 21 days after one-dose vaccination, the mortalities between the vaccinated group and the control group were 0% to 5% and 95% to 100%, respectively. In addition, the control group demonstrated a 95% to 100% re-isolation rate of the challenge strain in internal organs and the caecum, while in the vaccinated group only a 1% to 60% re-isolation rate was observed. In this study, we showed that adjusting the minimum vaccination age of the 9R vaccine to 4 weeks is acceptable considering the safety and efficacy of the vaccine.     

抗人Ⅳ型胶原蛋白单克隆抗体的制备和鉴定

用人Ⅳ型胶原蛋白免疫ＢＡＬＢ／ｃ小鼠，克隆了一组分泌抗人Ⅳ型胶原蛋白的杂交瘤细胞株，并且对其中的Ａ２、Ａ４、Ａ５、Ｂ３和Ｅ２进行了鉴定。其抗体亚类分别为ＩｇＧ１、ＩｇＧ２、ＩｇＧ２ｂ、ＩｇＧ２和ＩｇＧ１。其亲和力常数Ｋａｆｆ＝６．６×１０８Ｍ（－１）～６．０×１０（１０）Ｍ（－１）。它们均能特异性地识别人Ⅳ型胶原蛋白。     

Rapid, accurate genotyping of the common -alpha(4.2) thalassaemia deletion based on the use of denaturing HPLC

AIMS: To develop an alternative assay for specific genotyping of the -alpha(4.2) thalassaemia deletion based on the DNA sequence features surrounding the breakpoint. METHODS: The 5' and 3' ends of the breakpoint regions of the -alpha(4.2) allele and the normal homologous segments were sequenced in Chinese individuals. A sequence haplotype composed of four single nucleotide variations within the X2/X1 box of the -alpha(4.2) breakpoint region was found in all of the 10 Chinese -alpha(4.2) thalassaemia alleles studied. Based on these findings, a novel polymerase chain reaction (PCR)/denaturing high performance liquid chromatography (DHPLC) assay was developed for rapid genotyping of the -alpha(4.2) allele instead of traditional Southern blotting or Gap-PCR. This method involves amplification of the alpha globin target sequence encompassing these four polymorphic sites, followed by a partially denaturing HPLC analysis using the transgenomic WAVE DNA fragment analysis system. RESULTS: The three major genotypes (-alpha4.2/alphaalpha, -alpha(4.2)/--SEA, and alphaalpha/alphaalpha) could be distinguished through the characteristic chromatograms generated by the WAVE system. The accuracy of this technique was evaluated blindly, and the results were 100% (40 of 40) concordant with the genotypes previously characterised by Southern blotting or Gap-PCR. CONCLUSIONS: This study validates the PCR/DHPLC approach as a simple, rapid, highly accurate, and cost effective method, potentially adaptable for use in epidemiological surveys, genetic screening, and diagnosis of silent alpha+ thalassaemia and Hb H disease.