Objective To study the protective effects of estrogen on brain injured by cerebral ischemia-reperfusion in ovariectomized rats.Methods 30 d after bilateral ovariectomy,the benzestrofol 100 ?g/(kg?d)were intramusculari injected in to the models of ovariectomied rats for 14 d. Then the rat models of focal cerebral ischemia-reperfusion were made. The expressions of CD54 and TNF-? in brain tissue were detected by immunohistochemistry. The apoptotic cells were assayed by TUNEL,and the ultramicrostructural changes of neuron membrane was observed by electron microscope. Results Compared with the ischemia-reperfusion group and ovariectomized group,the expressions of CD54 and TNF-? of brain tissue in the estrogen group were significantly lower,and the apoptosis was reduced (all P

Objective To study the genetic polymorphisms of 11 Y chromosome specific STR loci in Guangzhou Han population. Method The DNA extracted from blood samples of unrelated individuals in Han population living in Guangzhou were amplified by PCR. The PCR products were analyzed by using PAGE. Results 3-5 alleles were detected in 11 Y - STR loci respectively in Han population in Guangzhou. The minimum GD value was 0.3037 (DYS434), while the maximum GD value was 0.8455 (DYS390) . Conclusion The 11 Y - specific STR loci are highly polymorphic and are suitable for personal identification and paternity testing.

AIM: To observe the effects of Salvia miltiorrhiza Bunge.f.alba. (Sal) on the mitochondrial ultra-structure, oxidative stress and apoptosis induced by ischemia injury in a rat model of focal cerebral ischemia and reperfusion.METHODS: The middle cerebral artery occlusion/reperfusion (MCAO/R) rat model was established by a modified Longa occlusion method. Adult male SD rats were randomly divided into control group, simple ischemia reperfusion group, Sal with ischemia reperfusion group and butylphthalide with ischemia reperfusion group. To study the protective effects of Sal and its mechanism, the intervention of Sal was given and the ultra-structure of mitochondria, functions of mitochondria under oxidative stress and the incidence of apoptosis of brain cells were determined.RESULTS: Many electron dense toxic granulation and vacuolus in mitochondria were observed in the rat brain of focal cerebral ischemia and reperfusion. Under the condition of ischemia and reperfusion, the mitochondria membrane was disaggregative, and the tubular cristae of mitochondrion disappeared. MDA content was obviously increased and the activity of glutathione peroxidase decreased significantly. The apoptosis of brain cells were observed in a great quantity. The changes of ultra-structure of mitochondria and the activity of GSH-Pxase were significantly improved by the treatment of Sal. Furthermore, treatment with Sal delayed the decrease of GSH-Pxase activity, and inhibited the increase in MDA content in brain tissue after ischemia and reperfusion. The incidence of apoptosis of brain cells was also decreased.CONCLUSION: Sal protects the brain tissue from ischemia injury.

AIM: To construct engineered E.coli strains which can express nattokinase with fibrinolysis activity using gene engineering technology. METHODS: The pro-nattokinase (pro-NK) gene was amplified by PCR and inserted into expression vector pET3c. The recombined plasmid pENK which expressed the fusion protein of pro-NK and 22 amino acid peptide was then transferred into lysogenic host strains BL21(DE3)pLysS - and BL21(DE3)pLysS +. Both SDS-PAGE and the fibrin plate assay were used to examine the expression and the activity of the target protein. RESULTS: SDS-PAGE assay showed the fused gene encoding 42 kD fusion protein was expressed in both expression strains pENK-(DE3)pLysS - and pENK-(DE3)pLysS +, and the fibrin plate assay indicated that the expression product had fibrinolysis activity. pENK-(DE3)pLysS - exhibited the basal expression of the target gene, while fusion protein was only induced by IPTG in pENK-(DE3)pLysS +. Basal expression of the fused toxic gene in pENK-(DE3)pLysS - led to bacteriolysis and hollow lawns. CONCLUSION: A pro-NK fusion protein with fibrinolysis activity is successfully expressed in E.coli , which lay a foundation for the exploitation of nattokinase.

AIM: To obtain a high and stable expression analog of human basic fibroblast growth factor by genetic engineering. METHODS: The cysteins 78 and 96 of natural hbFGF polypeptide was substituted with serines by means of site-directed mutagenesis. Using pET- 3c as vector, the mutated polynucleotide was cloned and then transferred into BL21 (DE3)plysS. After induction by IPTG, the analog was obtained and analyzed by SDS - PAGE. RESULTS: After purification the form of soluble mutant increased remarkably but the forms of dimmer and higher multimer were reduced greatly to no more than 8% of the total recombinant protein. By MTT assay, the analog showed the same biological activity. This new analog represented a desirable complementation for native hbFGF to develop pharmaceutical drug in clinical use. CONCLUSION: Substitution of certain amino acids of polypeptide without altering native protein' s bioactivity to get the analog is an effective means to increase stability of foreign protein and its solubility in E. coli.

Paeonia lactiflora Pall. has a long history of utilization, and is widely used in pharmaceutical, food and cosmetic industry. In this paper, the invention patents of traditional medicine P. lactiflora before 2014 were retrieved, and totally 18 192 patent families were obtained. And we formed the patent analysis report of medicine P. lactiflora based on a multi-angle analysis. Results show that, as to the patent number of medicine P. lactiflora, China is much more than any other country, and the applications mainly came from pharmaceutical enterprises. But the technologi-cal quality of patents and international protection ratio are low in our country. We need to strengthen in treatment of cardiovascular and liver disease. The patents mainly focused on the use of tonic health, which is compatible with Traditional Chinese Medicine. On the product development, dosage forms need to be enhanced. As to the compre-hensive utilization of resources, the flower and seed of P. lactiflora have relatively larger research space and value. This work will be helpful for researchers in deeply understanding the research achievements of medicine P. lactiflora, and provides the reference data for the future research.

Objective To explore the functional activation cerebral areas evoked by mechanical stimulation induced pruritus perception imitating physiological process.Methods Scratching the planta gently to induce pruritus with brush pen wool in a group of 16 healthy volunteers,scanning BOLD-fMRI serials with a block design on a 3.0 T MR machine,and identifying the functional cerebral areas evoked by pruritus with SPM8 software for analysis.Student's t test was used to compare the difference of incidences with P＜0.05 for statistical significance.Results Several brain regions were activated by pruritus stimulus.The strongest activation evoked by pruritus was found at the contralateral thalamus and paracentral lobule(t=5.26,5.23),the most activation volumes were found at the contralateral paracentral lobule,postcentral gyrus and prefrontal lobule(146,151 and 326 volumes).Different degrees of activation were discovered at bilateral insula,preeentral and postcentral gyrus,thalamus,lentiform nucleus and cingulated gyrus.Conclusion The postcentral gyrus,paracentral lobule,insula,precentral gyrus and frontal lobe were the functional activation cerebral areas of pruritus perception evoked by mechanical stimulation.

[Objective] To learn about the genetic diversity,we studied the five X-chromosomal STR (X-STR) loci in Guangdong Han Nationality Groups.[Methods] The five Loci (DXS6803,DXS981,DXS6809,DXS6789,and DXS7132) were amplified in a pentaplex PCR reaction.PCR products were analyzed using capillary electrophoresis and ABI prism 3100 Genetic Analyzer,with GeneMapper ID 3.1 Analysis Software.[Results] A total of 363 individuals (181 unrelated male and 182 unrelated female) from Guangdong Han population were tested,54 alleles were observed for these loci.Polymorphism information content is 0.6935 ～ 0.8177.Power of discrimination in females was 0.8976 ～ 0.9562.Mean exclusion chance for X-STR in standard trios with daughters was 0.7805 ～ 0.8467.[Conclusion] The five loci in the multiplex system provide high polymorphism information for forensic identification and paternity testing,particularly for difficult paternity deficiency cases.

AIM: To construct a recombinant hEGF-hbFGF(78-154aa)fusion protein, which not only has the heparin-binding ability, but also promotes the growth of the cells, and to express the fusion protein in E. coli expression system with high expression level.METHODS: hEGF gene was joined with 231 bp fragment coding hbFGF(78-154aa) and expressed in E. coli. The fusion protein was purified using affinity chromatography of heparin-Hyper D and analyzed with western blot. The pI value and the biological activity were both assayed.RESULTS: The fusion protein was expressed in a high expression level of about 30% of the total cell protein, as estimated by SDS-PAGE. Western analysis results showed that the antigenicity of fusion protein was similar to hEGF. Fusion protein could not only bind heparin but also promote the growth of 3T3 cell. The pI value of fusion protein was 5.2.CONCLUSION: The recombinant hEGF-hbFGF(78-154aa) fusion protein possessed the characteristics of both hEGF and hbFGF. This new-designed protein would become a good object for the research on the relationship between the structure and the function of the growth factor.

Objective To set up the electrochemical determination method for puerarin in Radix Puerariae.Methods The electrochemical behavior and content determination of puerarin were investigated by using coulomb array detector.Results In phosphate buffer with PH 4.32,puerarin showed good linearity in the range of 0.16 ng～ 162 ng.The regression equation was A=1444.7C+ 88.635,r=0.9998(n=7),the average recovery was 96.88 %,and RSD was 0.08 %.Conclusion This method is simple and rapid,and the result is accurate and reliable.It can be used for the determination of puerarin content and biological samples.