结核分枝杆菌酸抗性基因及其调控网络

病原菌在宿主细胞内的持留分子机理是目前研究的热点和难点。病原菌的抗酸能力与此密切相关。结核分枝杆菌感染导致的结核病仍然是全球公共卫生的重大威胁,这与结核分枝杆菌抗酸并在宿主巨噬细胞内持留有关。结核分枝杆菌抗酸主要通过调控质子进出、代谢调控胞内酸碱平衡和双组份信号系统调控。本文综述了结核分枝杆菌在酸胁迫下的整体调控网络,阐述了在酸性环境中结核分枝杆菌的具体调控机理,旨在为持留结核分枝杆菌的治疗提供新的全局性思路,寻找新的结核病防控靶标。

The Relationship of Methyl Jasmonate Enhanced Powdery Mildew Resistance in Wheat and the Expressions of 9 Disease Resistance Related Genes

[Objective] This study was carried out to determine the induction effect of jasmonic acid（JA）on powdery mildew resistance in wheat,the activation effect on the expressions of plant disease resistance related genes,and to investigate the relationship between the induced resistance and the gene expression patterns.[Method] Three powdery mildew susceptible cultivars of ＂Chinese Spring＂,＂Pumai 9＂ and ＂Zhoumai 18＂ typically representing different phenotypes in the field were employed.The powdery mildew was assessed by detached leaf assay,and real time quantitative RT-PCR was used to determine the expression patterns of 9 disease resistance related genes of PR1（PR1.1）,PR2（β,1-3 glucanase）,PR3（chitinase）,PR4（wheatwin1）,PR5（thaumatin-like protein）,PR9（TaPERO,peroxidase）,PR10,TaGLP2a（germin-like）and Ta-JA2（jasmonate-induced protein）in leaf of the three cultivars.[Result] MeJA application enhanced the powdery mildew resistances of ＂Chinese Spring＂,＂Pumai 9＂ and ＂Zhoumai 18＂.The induced powdery mildew resistance could be detected from 12 h to 96 h after MeJA treatment,and the peak value was at 24 h.Though there were differences between the three cultivars,MeJA significantly effect on the expressions of the 8 disease resistance related genes except TaGLP2a,and the peak values were at 12 h,24 h or 48 h after treatments.The strongest activation of MeJA was on PR9 and PR1 that their expressions could reach more than 100 times of the untreated samples.MeJA strongly activated PR2、PR4、PR5、PR3、PR10 and Ta-JA2,their expression could reach 10 to 70 times,and there was almost no activation effect on TaGLP2a.The induced powdery mildew resistance positively correlated with the induced expressions of the 8 disease related genes.[Conclusion] The induced powdery mildew resistance positively correlated with the induced expressions of the disease related genes.Jasmonate signalling plays a role in defence against Blumeria graminis f.sp.tritici.and future manipulation of this pathway may improve powdery mildew resistance in wheat.

PLASMA RESISTIN LEVELS AND SINGLE-NUCLEOTIDE POLYMORPHISMS IN RESISTIN GENE 5 FLANKING REGION IN PATIENTS WITH STROKE

Objective To analyze the role of resistin in insulin resistance (IR) through investigating the variation of plasma resistin levels and single-nucleotide polymorphisms (SNPs) in resistin gene 5’ flanking region in stroke patients.Methods In 103 atherothrombotic cerebral infarction (ACI) patients, 85 lacunar infarction (LI) patients, 70 intracerebral hemorrhage (ICH) patients, and 86 healthy controls, plasma resistin and insulin levels were measured by ELISA , SNPs in resistin gene 5’ flanking region were detected by PCR and direct DNA sequencing. The subjects’ body height and weight, the body mass index, quantitative insulin sensitivity check index (QUICKI), blood pressure, and the concentration of fasting plasma glucose, triglyceride, total cholesterol, creatinine, low-density lipoprotein, and high-density lipoprotein were also determined. Results QUICKI was significantly lower in the ACI and ICH patients (0.316±0.037 and 0.309±0.032, respectively) than that in the controls (0.342±0.043, P<0.001), while plasma resistin level was significantly higher in the ACI and ICH patients (6.36±3.79 and 7.15±4.27 ng/mL, respectively) than that in the controls (5.28±2.56 ng/mL, P<0.05), but such difference was not observed in the LI patients compared with controls. There was a statistically negative correlation between plasma resistin level with QUICKI (r=-0.228, P<0.001). The distributions of allele and genotype frequencies of resistin gene -420C>G and -537A>C SNPs were not significantly different among the different groups, and those SNPs were not correlated with other clinical and biochemical parameters.Conclusions Plasma resistin is associated with stroke by participating in the development of IR. The SNPs in resistin gene 5’ flanking region has no impact on the plasma resistin level.

ENDOGENOUS EXPRESSION AND HLA STABILIZATION ASSAY OF PLASMODIUM FALCIPARUM CTL EPITOPE MINIGENE IN HUMAN HLA- A2.1 AND HLA- B51 CELLS

To evaluate the Plasmodium falciparum CTL epitope vaccines in HLA class I allele specific human cell lines that have high frequency among Chinese population. Methods. Synthesized oligonucleotides encoding for P.f. CTL epitope genes, constructed eukaryotic expression plasmids, transfected the minigenes into HLA class I allele specific human cell lines and identified endogenous expressing of the minigenes by RT- PCR and HLA stabilization assay. Results. Two mini- genes encoding Plasmodium falciparum CTL epitopes were designed and cloned, respectively, into an eukaryotic expressing vector to form TR26 which was restricted to HLA- B51, SH6 which was restricted to HLA- A2.1, and TS, which had the two aforementioned mini- genes fused in tandem. All of these CTL epitope genes were transfected and endogenously expressed in respective cell lines containing appropriate HLA molecules. The obviously increased expressions of HLA class I molecules were detected in the transfected cell lines. It was demonstrated that the two discrete Plasmodium falciparum epitope genes were effectively processed and presented, and the close proximity of the two epitope genes in one chain as in mini- gene TS did not interfere with the processing and presenting of each epitope gene in corresponding cell line. Conclusion. A successful expression and presentation of multiple CTL epitope mini- gene in MHC class I allele specific human cell lines were demonstrated by an in vitro assay, which could be corresponding to the vaccination of CTL vaccines in people with different MHC I molecules. This work also suggested the possibility of constructing a multiple CTL epitope plasmodium falciparum DNA vaccine that could cover most of Chinese population.