Objective To screen the HBV SPⅡ promoter DNA-binding protein, and to investigate its potential role in the regulation of replication and expression of HBV DNA. Methods By using HBV SPⅡ biotinylated promoter DNA as a selective molecule, the T7 select human liver cDNA library was biopanned and the positive clones were selected. After screening, amplification of positive plaques was performed for inserted DNA fragment and then they were cloned into the pGEM-Teasy vector. Results Four positive plaques were chosen for DNA sequencing. The binding protein of HBV SPⅡ promoter was demonstrated as nicotinamide adenine dinucleotide (NADH) dehydrogenase 4 by BLAST. Conclusion The result suggests that this approach may provide a new tool for the study of replication and expression mechanism of HBV DNA.

Objective To screen the proteins binding with HBV X-promoter and to investigate their potential role in the regulation of replication and expression of HBV DNA. Methods By using HBV X biotinylated promoter DNA as the selective molecule, the T7 select human liver cDNA library was biopanned and positive clones were selected. After screening, positive plaques were performed to amplify for inserted DNA fragment and cloned into pGEM-Teasy vector. Thirty positive plaques were chosen for DNA sequencing. Results Five kinds of known and nine kinds of novel cDNA sequences were obtained. The 5 kinds of known ones are human serum albumin, NADH dehydrogenase subunit 4, prokininase, ubiquitin specific protease 10, and testis enhanced gene transcript. Conclusion The binding proteins of HBV Xp DNA were identified by phage display. The results suggest that this approach provides a new search tool for the study of replication and expression mechanism of HBV DNA.

AIM: To study the protocol and condition that induce bone marrow stromal cells (MSCs) to differentiate into neuron in vitro by baicalin, a kind of flavonoid isolated from an important medicinal plant Scutellariae Radix . METHODS: MSCs from adult rats were induced by baicalin in serum-free medium for 6 h, and postinduced for 6 d. The morphological changes of MSCs were evaluated by light microscope. The positive percentages of neuron-specific enolase (NSE), 200-kilodalton neurofilament (NF), glial fibrillary acidic protein (GFAP) and vimentin expression were measured by immunocytochemistry with ABC staining. Hoechst 33258 staining was used to measure the cell viability by fluorescence microscope. RESULTS: After induction for 6 d, MSCs displayed neuronal morphologies, such as pyramidal cell bodies and processes formed extensive networks. The positive percentages of NSE, NF, GFAP and vimentin protein expression were 70.5%?11.6%, 68.3%?13.4% ,

Molecular biology,one of the major branches of biochemistry,is quite abstract and abstruse for students.Fortunately with the help of multimedia,an effective teaching assistant method which enjoys great advantages,teachers can fully convey teaching ideas in a vivid picture.As a result,it will be much easier for students to follow teachers in class.

Cardiovascular disease, with high morbidity and mortality, has been threatening the health of human beings. Therefore, expecting to find a more effective therapeutic method, a plenty of researchers devote themselves to the study of the cardiovascular disease all the time. Since discovered on the heart, M3 receptor of muscarinic acetylcholine receptor (mAchR, M receptor) became a new starting point of the research of the cardiovascular disease. With more and more investigation, many people found that M3 receptor could protect the heart from kinds of cardiovascular disease, which may make it a new hopeful therapeutic point. So, expecting to give support to the reference and encouragement for the study of disease related to M3 receptor in future, this review expounds M3 receptor on the heart from the main following aspects: the effect on the heart, the influence on the cardiovascular disease and the mechanism of M3 receptor involved.

Objective Frontal resting EEG asymmetry was used to evaluate the gender effect of emotion regulation.Methods The resting EEG was recorded in 30 healthy volunteers (19-26 years old) by the guide 40 amplifier,and off-line analysis on EEG data were conducted by scan4.3 software.The statistical analysis were performed using SPSS 17.0.Results In normal human,frontal resting EEG alpha power was statistically significant(F=4.918,P =0.035).Asymmetry and gender interaction group (F =0.668,P =0.421),electrode point and gender interaction group (F =0.283,P =0.756),electrode point and asymmetry interaction group (F =1.418,P =0.260),electrode point,asymmetry and gender interaction group (F =1.164,P =0.327) had no statistical significance (P ＞ 0.05).The comparison of male and female asymmetry degree of each electrode point (F4/F3 group t =0.465,F8/F7 groupt=0.809,FP2/FP1 group t=1.542) had no statistical significance (P＞0.05).Conclusion Normal human brain hemispheres exist asymmetry.The asymmetry does not exists gender effect.In non-task state,frontal resting EEG alpha power asymmetry can not be recognized as the indicator of emotion regulation ability.

AIM:To construct a eukaryotic expression vector containing pancreatic duodenal homebox-1(PDX-1)and to elevate the expression efficiency of exogenous gene in rat bone marrow mesenchymal stem cells(MSCs).METHODS:Recombinant vector containing PDX-1 was constructed.Flow cytometry was used to identify the cell cycle of bone marrow mesenchymal stem cells(MSCs)cultured in vitro.Recombinant vector containing PDX-1 was transfected into bone marrow MSCs using superfect in medium.After being selected by G418,RT-PCR and Western blotting were used to investigate the expression of PDX-1 in MSCs.RESULTS:Restricted enzyme analysis and sequencing showed that PDX-1 gene segment was consistent with that in GenBank.Flow cytometry showed that there were about 85.9% cells at the cell cycle of G_0/G_1.The whole cells transfected emitted green fluorescence under flow cytometry.The efficiency of transfection was above 40%.RT-PCR and Western blotting demonstrated that there was expression of PDX-1 in transfected bone marrow MSCs.CONCLUSION:Recombinant vector containing PDX-1 was constructed successfully.Superfect mediated expression of exogenous gene in bone marrow MSCs in a high efficiency,and bone marrow MSCs containing exogenous gene are an ideal cells for gene therapy.

Objective To investigate a new method of a long term culture for rat islets in vitro. Methods Rat islets marked by green fluorescent protein (GFP) were cocultured with Sertoli cell for 20 weeks. Histological studies were performed on islet group and coculture group in 1w, 3w, 10w, 15w, 20w of culture by light, fluorescent and electron microscopy. Insulin released was measured by radioimmunoassay. Results In islet group, islet viability and the number of insulin positive cells were significantly decreased after 3w of culture, cumulative quantities of insulin for 24 hours and the stimulation index also fell rapidly under this condition, meanwhile the ultrastructure of islets was destroyed. However, under coculture condition, culture time of islets was prolonged in vitro, islet viability and the number of insulin positive cells were significantly increased, cumulative quantities of insulin for 24 hours and the stimulation index maintained at high level, and the ultrastructure of islets remained normal even after 20w of culture. Conclusion Coculture of rat islets with Sertoli cells may promote islet growth and prolong culture time, and it is a new method of a long term culture of islets in vitro.

AIM: To investigate whether grafting neural stem cells (NSCs) improves the impaired cognitive deficits and spatial recognition after ischemic-hypoxic brain damage (HIBD) in neonatal rats. METHODS: Non-immunosuppressed 7-day-old SD rats were used as research subject and randomly divided into 3 groups: (1) sham group (n=10); (2) HIBD group (n=11); (3) transplant group (n=13). (2) and (3) were anesthetized and subjected to a hypoxic/ischemic injury obtained by combination of left carotid ligation and exposure to 8% oxygen for 2 h. At 3 days post injury, hypoxic-ischemic brain damaged animals were re-anesthetized and randomized to receive stereotactic injection of NSCs prelabeling with BrdU or control media into the hippocampus in the ipsilateral hemisphere. Cognitive (i.e., learning) deficits were assessed at 2 to 4 weeks after transplantation. At the end of the behavioral tests, the animals were killed and evaluated for NSC survival and histopathological analysis. RESULTS: Transplant group showed significantly improved cognitive function in selected tests as compared with HIBD group during the 4-week observation period. They took less time than HIBD group in finding the 3 arms baited with water and had a decreased number of working and reference memory errors in radial maze acquisition tests. Histological analysis showed that transplanted NSCs attenuated CA1 cell loss after HIBD, and NSCs survived for as long as 4 weeks after transplantation and were detected in the hippocampus. CONCLUSION: These data suggest that transplanted NSCs attenuate brain damage and cognitive dysfunction after hypoxic-ischemic brain damage. This approach warrants continued investigation in light of potential therapeutic uses.