Experimental study on how brain-dead state affects the heart structure and function of Ba-Ma mini pigs and the mechanism

AIM:To investigate how brain-dead state affects the heart structure and function and the effect of PKC-α in BA-Ma mini pigs.METHODS:Ten Ba-Ma mini pigs were randomized into 2 groups: brain-dead group (n=5),and control group (n=5). The brain-dead model was made by increasing intracranial pressure,while the control group was maintained anesthesia for 24 h. The concentrations of cTnT,TNF-α,IL-1β and IL-6 in serum were determined at 6,12 and 24 h after brain death. At 24 h,heart tissues were observed by HE staining and electron microscope. The expression of PKC-α was detected by immunohistochemistry and RT-PCR.RESULTS:(1) Histological changes of myocardium: flaky bleeding under endocardium and dissolution of myocardium were found in optical microscope. In electron microscope dropsical mitochondria and confluent muscle fiber were found. (2) Changes of serum cTnT: serum cTnT for brain-dead group began to increase gradually since 6 h,and were significantly higher at each time point than those in control group (P<0.05). (3) Changes of inflammatory factors: IL-1β,IL-6,and TNF-α in brain-dead group began to increase gradually since 6 h,and were significantly higher at each time point than those in control group (P<0.05). (4) Changes of PKC-α expression: PKC-α mRNA and protein expressions in brain-dead group increased significantly at 24 h (P<0.05).CONCLUSION:Brain death may evoke heart structure and functional injury,and increase the levels of inflammatory factors and PKC-α. The activation of PKC-α may participate in the process of heart injury.     

Nucleostemin gene expression in human breast tumor tissues and its relation with clinical pathology

AIM: To study the expression of nucleostemin (NS) gene in human breast tumor tissues and the relations of NS gene expression level with histological grades,histological types and TNM stages of the tumor.METHODS: Total RNA was isolated from human breast tumor tissue.The methods of electrophoresis and RT-PCR were used in measuring NS gene expression level,and the relations of NS gene expression level with histological grades,histological types and TNM stages of the tumor were analyzed.RESULTS: The results indicated that there was no NS gene expression detected in normal breast tissues,and NS gene expression in malignant breast tumor tissues (P<0.01) was higher than that in the benign breast tumor tissues.The higher histological grades of the breast cancer showed the stronger NS gene expression (P<0.01),the higher TNM stages of the breast cancer showed the stronger NS gene expression (P<0.01),and the level of NS gene expression had not correlation with the histological types (P>0.05).CONCLUSION: It is suggested that there is no relation of NS gene expression level with histological types of the breast cancer,but there is a marked correlation of NS gene expression level with the histological grades and TNM stages.     

Sequence analysis and prokaryotic expression system construction of PIA genes isolated from Neisseria gonorrhoeae

AIM: To analyze the nucleotide and putative amino acid sequences of PIA genes isolated from N.gonorrhoeae and to construct the prokaryotic expression system of PIA gene.METHODS: The entire PIA genes from 9 strains of N.gonorrhoeae were amplified by using high fidelity PCR.The target amplification fragments were sequenced after T-A cloning.Homology comparison of the nucleotide and putative amino acid sequences of PIA genes from the isolates with the reported sequences in GenBank was then performed.A prokaryotic expression system of PIA gene was constructed.Different dosages of IPTG were applied to induce the expression of the target recombinant protein (rPIA) and 10% SDS-PAGE plus Bio-Rad Agarose Image Analysor was used to determine the expression level of rPIA.rPIA was extracted using Ni-NTA affinity chromatography and the purified effect was detected by SDS-PAGE.RESULTS: In comparison with the reported PIA gene sequences (GenBank No: L19962),the homologies of nucleotide and putative amino acid sequences of PIA genes from the isolates were 99.6%-100% and 99.1%-100%,respectively,which indicated that all the isolates were belonging to serovars IA6.Output of rPIA was as high as 50.1% of the total bacterial proteins.The purified rPIA only showed a single target protein fragment in gel.CONCLUSION: Serovar IA6 is dominant in the local N.gonorrhoeae isolates and sequences of the encoding gene are relatively conserved.The constructed prokaryotic expression system is able to express rPIA with high efficiency,which may lay a foundation for further development of serological detection kit and vaccine of N.gonorrhoeae.     

万寿菊细菌性叶斑病药剂防治试验

通过室内抑菌筛选和田间小区及生产试验,从当前防治细菌性病害的药剂中,筛选出对万寿菊细菌性叶斑病防治效果较好的3种药剂及最佳使用浓度:90%新植霉素150mg/L,细菌清200 mg/L和72%农用链霉素150 mg/L,其防效分别为73.6%,72.7%和70.7%,且药效稳定,增产效果显著.     

Effect of advanced glycation end products on inflammation in cultured cardiomyocytes

AIM: To investigate the effect of advanced glycation end products on inflammation in cultured cardiomyocytes.METHODS: Primary cardiomyocytes were isolated from Sprague-Dawley neonatal (1 to 2 days old) rats ventricles.The insulin resistant cardiomyocyte model was established.Neonatal rat ventricular myocytes were exposed to AGEs for 24 hours.TNF-α mRNA and PPAR-γ mRNA expressions were determined by RT-PCR.Activation of NF-κB in the cells was examined by using immunocytochemistry.The ultrastructure of the cells was detected by transmission electron microscope.RESULTS: The exprssion of TNF-α mRNA and the activation of NF-κB increased,the expression of PPAR-γ mRNA decreased compared with control group (P<0.05).The differences among different AGE-BSA groups were significant (P<0.05).The numbers of chondriosome and smooth endoplasmic reticulum increased.CONCLUSION: AGEs significantly increase TNF-α mRNA expression and NF-κB activation,and restrain the expression of PPAR-γ mRNA.These data suggest that AGEs play an important role in the onset of diabetic cardiomyopathy.     

Regulation of estrogen on the expression of KLK6 mRNA and protein in human breast cancer cell line MCF-7

AIM:To investigate the effects of estrogen and tamoxifen on the expression of KLK6 mRNA and protein (hK6) in human breast cancer cell line MCF-7. METHODS:MCF-7 cells were incubated with 17-βE2 and tamoxifen at different concentrations for 72 hours, respectively. The expression levels of kallikrein 6 (KLK6) mRNA and protein were evaluated by fluorescence quantitative RT-PCR and flow cytometry, respectively. RESULTS:Compared with ethanol control, KLK6 mRNA expression levels were significantly decreased when 17-βE2 was added at concentrations of 10^-10 and 10^-8 mol/L (P<0.01). No statistical change was observed when 17-βE2 was at 10^-12 mol/L (P>0.05). Flow cytometry showed the same results. The average fluorescence intensity (AFI) that represents the level of hK6 was decreased after incubated with 17-βE2 (P<0.01). After incubation with tamoxifen, the levels of KLK6 mRNA and hK6 were increased (P<0.01). CONCLUSION:Estrogen down-regulates the expression levels of KLK6 mRNA and protein (hK6), while tamoxifen has an opposite effect.     

县域农业可持续发展的生态安全评价——以吉林省通榆县为例

以吉林省通榆县为例,构建了县域农业可持续发展的生态安全评价指标体系,采用均方差法确定了评价指标的权重,对研究区1985、1990、1995、1998、2001、2004年6个时段的农业生态安全进行了评价.从评价结果可以看出,近20年,通榆县农业生态安全呈现下降态势,安全等级处于Ⅲ～Ⅴ级,安全状态从敏感态下降到风险态.1998年是研究区农业生态安全变化的转折点,从1998年起,农业生态安全有所回升.在分析变化原因的基础上,提出农业可持续发展的生态安全建设对策.     

渭河下游水量变化的防洪响应

在对渭河下游的来水来沙量进行分析的基础上,结合水槽试验结果和实测资料,从机理上探讨渭河下游呈现出洪水历时延长、滩地高程不降、水位处于逐渐升高的态势、边界条件劣势发展的不利防洪响应,认为其主要原因是产生贴边淤积的环境良好.据此,针对该环境所引起的防洪形势严峻问题,提出了治理对策.