Abstract Objective:To better understand the regulatory mechanism of Pax-5 gene expression on B-cell differentation and development.Methods:The isolation and genomic colning of the 5'-flanking region of Pax-5 gene were carried out according to established protocols: molecu-lar cloning and subcloning The nucleotide sequences were determined by a double-stranded dideoxy sequencing method. Sequence analysis wasperformed on line programs. Results:Analysis of tha total sequence(6 671 bp) shows the proximal promoter include 3 CAT boxes, 1SP1 and 1Ebox. However, there was no consensus sequence for a TATA box in the 5'-flanking region. Putative regulatory sites of further upstream in thesequence revealed 6LMO2 COM, 5NFAT, 2LPOLYA-B, 3GATA1, 2AP4, 10MZF1, 1ets1B, 1GATA3, 1NKX-25, 2RORA1, 1LYF1, 2Ikaros2,2TCF11 , 1GATA-C and 1FREAC7. Conclusion: The 5'-flanking regionn of human Pax-5 gene exon1B could be involved in regulating the expres-sion of human Pax5 and B-cell differentiation and development.

Objective To construct the lamino acid transporter 1 eukaryotic expression vector of C 57 mouse and to express the gene inNeuro-2atumor cells,and explore the effect of LAT1on proliferation and apoptosis of Neuro-2a cell. Methods The full-length LAT1 cDNA was synthesized by RT-PCR and cloned into pcDNA3.1vector to construct recombinant plasmid.The constructed pcDNA3.1-LAT1vector was verified by Enzyme digestion and sequencing and then transfected intomurine Neuro-2acellsby liposome.The transfected cells were selected with G418 and stably expressed strain was constructed .The expression of LAT1 was detected by RT-PCR and western blot .Proliferation was analyzed by MTT , cell cycle and apoptosis were detected by flow cytometric analysis .Results The full-length LAT1 cDNA was amplified successfully and pcDNA3.1-LAT1eukaryoticvector was constructed successfully .Enzyme digestion and sequencing confirmed the sequence was correct .Neuro-2acells were transfected and Stably expressed strain was constructed successfully.MTT showed that the group of transfected restructuring plasmid could significantly affect Neuro -2a cell proliferation more than the control groups ( P <0.05 ) .From the flowcytometric analysis , LAT1 could promote cell proliferation and inhibit Neuro-2a cell apoptosis.Conclusion LAT1 can express successfully inNeuro-2acells which were transfected with recombinantpcDNA3.1-LAT1plasmid.LAT1 in Neuro-2a cells can promote cell proliferation and inhibit the cell apoptosis which provides a basis for the study of LAT 1.That lays the foundation for studying biological effects of LAT 1.

Objective To detect the expression changes and location of the mouse developmental regulation brain protein(Dbn1)in the developmental mouse brain.Methods Monoclonal antibodies against drebrin protein were used to assess Dbn1 by immunohistochemistry and Western blot.The paternal expression of Dbn1 in developmental mouse brain(E14,P1,P7 and adult)was initially investigated by Western blot.Dbn1 was shown at various developmental stages(E14,P1 and P7)as well as in adult in different brain area of developmental mouse brain by immunohistochemical.Results Dbn1 protein was detected in developmental brain but a little in adult brain by Western blot,high at E14,decreased at P1,gradually increased at P7 and lowest in adult.Immunohistochemistry confirmed as follows:Dbn1 expressed mostly in cortex,hippocampus and ependyma areas at E14,and the positive signal was distributed at cells border;The expression of Dbn1 was decreased at P1,mainly distributed at the verge of cells or dendrites;The peak expression of Dbn1 appeared at P7,Dbn1 located at nucleus of hippocampus and cortex,and the positive signal located within cytoplasm and dendrites;Only a little positive Dbn1 cells were found in adult mouse brain.Conclusion Dnb1 may be involved in regulating the differentiation and migration of neurons during the development of mouse brain.

It is important and necessary for the teaching process in histology and embryology integrated by psychological quality.The psychological quality of teachers can be improved by professional training and by themselves.Teachers should teach everyone differently according to the different psychological character of medical students who are born after 1990.Teachers can improve psychological quality of medical students in teaching process including the discussion,visiting the embryo sample and the second class.

Due to the particularity of marine ecological environment, many marine lives have developed and accumulated a large amount of biological molecules with special chemical structures and physiological activities, representing an important resource for the development of marine drugs. Generally, the marine antibacterial proteins are mainly identified from the marine microorganisms, invertebrates and fish.In this review, the progresses on marine antibacterial peptides and antimicrobial activity proteins are briefly summarized.

Objective:Study on characteristics of two synthesizd peptides based on CSFV E2 protein. Methods:B cell epitopes of CSFV E2 antigen were predicted using accessibility and flexibility schemes, associated with antigenicity , secondary structure and multiple sites prediction. Two antigen peptides (Pep1 and Pep2) have been designed and synthesized and their reactivety were detected with 8 McAbs and antiserum against mE2 protein, then the peptides were conjugated with BSA and immunized rabbits respectively. Results:Both Pep1 and Pep2 could react with antiserum and McAb A11, Pep2 could interact with McAbD5 and McAbD8. Only Pep1 BSA conjugate can stimulate high level and specific antibodies.Conclusion: The peptide1 has good antigenicity.

Objective To isolate and identify pancreatic stem cells of Kunming mouse and to observe the differentiation potency of those cells. Methods Pancreas of postnatal Kunming mice were digested by enzymes to isolate pancreatic stem cells. The potency of the cell differentiation was then identified with special marker of cytokeratin 19(CK 19), insulin and glucagons by immunocytochemical staining. Results Few epithelioid cells could be found to survive at the beginning of isolation, but when medium was replaced by that with growth factor, the cells proliferated quickly in fusiform shape and formed colony gradually. The cells were CK 19 immunoreactive positive after transfer of culture. After differentiation induced by high glucose, the cells formed the pancreatic islet like structures. Immunocytochemical staining showed that part of the cells of pancreatic islet like structures were insulin immunoreactive positive and few of the cells were glucagon immunoreactive positive. Conclusion Pancreatic stem cells of Kumming mouse can proliferate when cultured in vitro and have the potency of differentiation into ? and? cells of pancreatic islet.

OBJECTIVE: To study the role of nuclear factor-kappa B (NF-kappaB) and cycloxygenase-2 (COX-2) in the onset of phlegm obstruction due to lung-deficiency in rats and the therapeutic mechanism of Huatan Recipe. METHODS: Twenty-four SD rats were randomly divided into normal group, model group and treatment group, with 8 rats in each group. The rats in the model group and treatment group were exposed to sulfur dioxide and cold wind to establish the rat model of phlegm obstruction due to lung-deficiency, and the rats in the treatment group were also treated with Huatan Recipe, a compound traditional Chinese medicine. The expression of NF-kappaB in the bronchial epithelial cells of the rats was tested with the method of immunohistochemistry, and the COX-2 mRNA in the lung tissues of the rats was measured by using reverse transcription-polymerase chain reaction. RESULTS: The expressions of NF-kappaB and COX-2 mRNA in rats of the model group were higher than those of the normal group (P<0.01), and the expressions of NF-kappaB and COX-2 mRNA in rats of the treatment group were obviously lower than those of the model group (P<0.01). CONCLUSION: The NF-kappaB and COX-2 play an important role in the onset of phlegm obstruction in rats. Huatan Recipe may prevent the development of phlegm obstruction by down-regulating the expressions of NF-kappaB and COX-2 mRNA.

ObjectiveTo investigate the effect of Tongxinluo by the dynamic changes of the expression of matrix metalloproteinase-9(MMP-9) on local cerebral ischemic-reperfusion injury in rats.MethodsMale Wistar rats were randomly divided into sham-operated control group, normal saline control groups (24-hour and 5-day) and Tongxinluo treatment groups(24-hour and 5-day). Animals in the latter four groups were subjected to focal ischemia by filament occlusion of the middle cerebral artery for 90 min. At the different end point, the expression of MMP-9 were assayed using immunohistochemistry staining method. ResultsCompared with the control groups, the expressions of MMP-9 were significantly decreased in both 24-hour and 5-day Tongxinluo treatment groups(P<0.05).ConclusionThe protective effects of Tongxinluo on cerebral ischemia reperfusion injury may be mediated by pharmacological inhibition of MMP-9 activity.The protective effects of Tongxinluo on cerebral ischemia reperfusion injury may be mediated by pharmacological inhibition of MMP-9 activity.

[ ABSTRACT] AIM:To observe the changes of perilipin and adipose differentiation-related protein ( ADRP) du-ring the development of diabetes mellitus and to explore the effect of perilipin and ADRP on abnormal glucose metabolism with non-alcoholic fatty liver disease ( NAFLD) .METHODS:The rat model of impaired glucose tolerance ( IGT) was in-duced by feeding high-fat diet, and the type 2 diabetes mellitus (T2DM) model was induced by feeding high-fat diet for 4 weeks and intraperitoneally injecting streptozotocin.The morphological change of the liver tissue was observed under optical microscope.The serum contents of perilipin and ADRP were measured by ELISA.The mRNA expression of perilipin and ADRP in the liver tissues was detected by real-time PCR.The protein expression of ADRP in the liver tissues was deter-mined by Western blotting.RESULTS:HE staining showed steatosis in the liver of the rats in IGT group was more serious than that in T2DM group.The biochemical and the pathological processes of rat models were consistent with the clinical feature of related diseases.The serum content of perilipin had no difference among various groups.The mRNA expression of perilipin in IGT group and T2DM group was significantly higher than that in control group.Compared with IGT group, the mRNA expression of perilipin in T2DM group was significantly increased.The serum content of ADRP in T2DM group was significantly lower than that in control group.The mRNA and protein expression of ADRP in model groups was signifi-cantly lower than that in control group.Compared with IGT group, the mRNA expression of ADRP in T2DM group was sig-nificantly reduced.CONCLUSION: The serum content of ADRP plays a role in the development and progression of T2DM.It is negatively correlated with HOMA-IR.NAFLD occurs during progression of abnormal glucose metabolism in-duced by feeding high-fat diet.The development of abnormal glucose metabolism with NAFLD is probably related to the in-creased expression of perilipin and the reduced expression of ADRP.