The study of traditional Chinese medicine and its complex prescription gradually adopted the methods of serum pharmacology, which has become a new researching thought and a gained more attention. In this article, the background of relevant researches, methodological discussions, analysis of the advantages and problems of serum pharmacology were reviewed. The serum pharmacology, for its unique value, will certainly has a brilliant prospect in its application to the research of traditional Chinese medicine and its complex prescription.

The goal of this work is to understand the difference between vertical structure and horizontal structure on data management. We take adverse event report as an example to find out the relationship between the storage space and the factors related to the trail. From the study we find out the difference of storage space between horizontal structure and vertical structure goes up with the increase of the storage space of each adverse event record and the number of adverse event allowed to record in the horizontal structure and goes down with the increase of the happening rate and the average times of the adverse event. When using the most widely used study design, vertical structure always takes less storage space than the horizontal structure. This vertical structure database is more suitable for the data management than the horizontal structure database.

Objective To explore the changes of cytokines in myasthenia gravis children and the effect of glucocorticoid on the serum levels of them.Methods Forty cases of myasthenia gravis children were the observation group which was divided to group one (pre-glucocorticoid therapy)and group two (post-glucocorticoid therapy),and 20 cases of healthy children in the same period as the normal control group.The serum levels of cytokines INF-γ,TGF-β1,IL-10 and IL-18 of the observation group one and two and the normal control group were detected by ELISA and were compared between the observation group one and the normal control group and the observation group one and two.Results The serum levels of cytokine INF-γand IL-18 were higher and IL-10 and TGF-β1 were lower in the observation group than in the normal control group,the differences were statistically significant (t =4.45 ~16.72,P <0.01 ).Significant difference of INF-γ,TGF-β1 and IL-18 was found between the observation group one and two (t =8.12 ~10.68,P <0.01)and the sera level of IL-10 had no significant difference (P >0.05).Conclusions Cytokines INF-γ,TGF-β1,IL-10 and IL-18 are involved and probably play different roles in the pathogene-sis of myasthenia gravis in children;Glucocorticoid could affect the secretion of cytokines IFN-γ,TGF-β1 and IL-18 of myasthenia gravis children,which would ultimately to achieve the aim of interfering and con-trolling the clinical symptom of myasthenia gravis in children.

Objective The purpose of this study was to evaluate the disinfection effectiveness and properties of acrylic resin via microwave irradiation. Methods Forty acrylic resin base dentures fabricated in a standardized procedure were chosen and divided into Group A,B,C and D randomly. Group A,B and C were immersed in 200ml distilled water and submitted to microwave irradiation at 700W for 3,4,5 minutes individually. Group D was used as positive control. Bacteria specimens from each group were got for culture and numbers were calculated. Then the four groups were tested for the flexural strength and impact strength by universal test machine before and after microwave irradiation sterilization. Results The germicidal ratio of Group B was more than 90% ,while the ratio of group C was 100%. There was no significant difference in flexural strength and impact strength between group C and group D. Conclusions Microwave irradiation for 5 minutes at 700W produced sterilization of dentures contaminated with all bacteria whereas the denture strength is not affected. Microwave irradiation at 700W solution for 5 minutes was effective to sterilize acrylic resin base dentures.

Objective To investigate the proof of fetal cells passing through the placental barrier into the maternal peripheral blood to provide laboratory data for the non invasive prenatal gene diagnosis of genetic diseases. Methods A total of 22 samples of placental tissues delivered(male fetus: 12, female fetus: 11) were divided into two groups for parallel section. HE staining was used to find the distribution of fetal cells in chorionic villi. In situ hybridization (ISH) technique with SRY DNA probes was used to identify the existence of fetal cells in placental villi, particularly in intervillous space. Results Light microscope examination revealed that there were fetal cells that passed through the capillary endothelium of villi and trophoblast basement membrane in the placental tissue sections of the 22 samples. ISH with SRY DNA probe also revealed that there were positive signals in the capillary of villi, at the edge of trophoblast basement membrane and in intervillous space in the placental tissue sections of the 12 placentas, but no signals were found in 10 female placentas. Conclusion This study demonstrates that the distribution of the fetal cells in the chorionic villi and intervillous space could be identified. The detection of fetal DNA in maternal circulation is one of the candidate approaches for non invasive prenatal gene diagnosis.

Kruppel-like factor 6 (KLF6) was reported as tumor suppressor in multiple cancers. However, loss of chromosomal locus spanning KLF6 is relatively infrequent in previous published studies. To explore the role of KLF6 in hepatocellular carcinoma (HCC), we examined the gene for expression change, loss of heterozygosity (LOH) and mutation in 26 HCC samples. The expression levels of KLF6 were significantly down-regulated in HCCs, as detected by qRT-PCR. LOH occurred in 11 (52%) of 21 tumors, and all the samples with LOH showed KLF6 down-regulation. The mutational frequency was 24%, and sequence changes located in activation domain of KLF6. Furthermore, MTT assay showed a significant antiproliferative effect of the wt KLF6 transfected in HepG2 hepatoblastoma cells. Fluorescence-activated cell sorting analysis revealed that KLF6 could induce apoptosis. These findings indicate that deregulation of KLF6, together with genetic abnormalities of allelic imbalance and mutations, may play a role in HCC pathogenesis.

Objective To explore the potential mechanism of abnormal behavior resulted from maternal separation in neo-natal period in rat. Methods Neonatal rats were equally and randomly divided into maternal separation group and control group. The rats in maternal separation group were separated from the dam for 3h per day on postnatal days (PND) 2 to 21, nothing was done to the rats in the control group. The brain tissues were taken out after being killed on PND 7, 14, and 21. The expressions of Caveolin-l, BDNF and GFAP in hippocampal formation were detected by immunohistochemistry. Semiquantitative assessment of immunohistochemical images was performed by Image-Pro Plus software. Results Compared with control groups, the expres-sion of Caveolin-l on PND 7 had no signiifcant change, while BDNF and GFAP were signiifcantly increased in maternal separa-tion group (P<0.05). On PND 14 and 21, the expressions of Caveolin-l, BDNF and GFAP were signiifcantly decreased in maternal separation group (P<0.05). Conclusions Decreased expressions of Caveolin-l, BDNF and GFAP caused by maternal separation in neonatal period may be associated with abnormal behaviors in adulthood in rat.

Objective To express and purify the recombinant nucleoprotein fragments of hemor-rhagic fever with renal syndrome ( HFRS) virus and to evaluate their diagnostic efficacy by using array-ELISA technology .Methods The target genes encoding nucleoprotein fragments of HFRS virus were amplified by PCR, and then inserted into prokaryotic expression vectors to construct the recombinant plasmids of pET -32a (+)/Pn and pET-32a(+)/Pc.The plasmids were transformed into E.coli BL21 ( DE3) to induce the ex-pression of nucleoprotein fragments by IPTG and the expressed products were purified by affinity chromatog -raphy using Ni-NTA agarose.The specificity and sensitivity of the recombinant antigens were evaluated by the assay of array-ELISA using commercial colloidal gold assay kit as a comparison .Results The recombi-nant nucleoprotein fragments of HFRS virus were correctly expressed in E.coli and highly purified by affinity chromatography .Array-ELISA showed that 13 of 16 suspected serum samples were positive by using the His-Pn protein as diagnostic antigen , consistency with the commercial colloidal gold assay kit reaching 94%. Conclusion The recombinant His-Pn protein expressed in E.coli cells could be used for specific serodiag-nosis of HFRS virus as its high antigenicity and sensitivity .The array-ELISA is an effective assay for the de-tection of virus at protein level .

Objective To express and purify Japanese encephalitis virus ( JEV) EDⅢprotein and evaluate the possibility of using it as a candidate antigen in JEV diagnostic kit .Methods PCR primers spe-cific for the gene encoding JEV EDⅢprotein were designed and used to amplify the gene fragment by RT-PCR.The cloned gene fragment was then inserted into pET-30a (+) to construct the recombinant expression plasmid.The transformed E.coli BL21 carrying expression plasmid were induced by IPTG to express JEV EDⅢ protein.The expressed JEV EDⅢprotein and a control antigen of tick-borne encephalitis virus protein were deposited in small spots to set up ELISA microarray .The serum samples from patients with Japanese encephalitis and healthy people were detected by Array-ELISA.The results obtained by Array-ELISA were compared with those by using indirect immunofluorescence assay .Results The gene fragment encoding JEV EDⅢprotein was successfully cloned and expressed in E.coli BL21.The recombinant protein could be used in Array-ELISA assay for the detection of serum samples from patients with Japanese encephalitis and healthy subjects .The results were consistent with those by using indirect immunofluorescence assay.Conclusion The recombinant JEV EDⅢprotein can be used as a candidate antigen for the diagnosis of JEV infection .

Objectives To explore the role of PKCβ/p66Shc oxidative stress signaling pathway in hyperoxia-induced apoptosis of alveolar epithelial cells A549. Methods A549 cells were cultured in vitro and divided randomly into control (incubated with 5%CO2), hyperoxia group (exposed to a mixture of 900 ml/L O2 and 50 ml/L CO2 at speed of 3 L/min for 10 mins, then cultured in a closed environment) and LY333531 group (treated with 10μmol/L of PKCβinhibitor LY333531 for 24h then induced with hyperoxia for 10 mins). The cellular morphology was observed under inverted microscope at 12, 24 and 48 h of treatment. The cell apoptosis was detected by lfow cytometry. Expression of PKCβ/Pin1/p66Shc/p66Shc-Ser36 were detected by immunohistochemistry after 24 h of treatment. Results Comparing to the control group, the cellular morphology of A549 in the hyperoxia group changed to spherical shapes and space between cells increased, the living cell count decreased and suspension cell increased. The living cell count in LY333531 group increased and suspension cell decreased than those in hyperoxia group but not reach the levels of the control group. The apoptosis rate of A549 cells and the expression of PKCβ/Pin1/p66Shc/p66Shc-Ser36 at 24 h were signiifcantly increased in the hyperoxia group than those in the control group, while the apoptosis rate and the expression of PKCβ/Pin1/p66Shc/p66Shc-Ser36 were greatly decreased in the LY333531 group than those in the hyperoxia group (all P<0.01). Conclusions The expression of PKCβin A549 cells can be increased by the hyper-oxia induction but reduced by LY333531, and then the expressions of Pin1, p66Shc and p66Shc-Ser36 are reduced. Thus the re-duced apoptosis of A549 cells relieve the cell injury induced by hyperoxia.