Objective It has been unclear how osteopontin, one of metastasis-associated genes, could be regulated in HepG2 cells. The aim of this study was to investigate the effect of protein kinase B(Akt) on the expression of osteopontin in HepG2 cells, and to explore the relationship between Akt and osteopontin expression. Methods HepG2 cells were transfected with constitutively active Akt and dominant negative Akt by lipofectmine, and transfectants were confirmed using Western blot for Akt. Osteopontin expression was detected by Northern blot and Western blot. Results HepG2 cells were successfully transfected with Akt genes including constitutively active Akt and dominant negative Akt, and overexpression of exozegenes Akt could be detected in HepG2 cells by Western blot. Using Northern blot and Western blot, we found that Akt gene regulated osteopontin expression in RNA level and protein level. In serum-free condition, HepG2 cells constitutively expressed low levels of osteopontin. Transfection of constitutively active Akt increased osteopontin mRNA and protein expression. Transfection of dominant negative Akt decreased osteopontin expression.Conclusions The fact that osteopontin gene expression can be regulated by Akt indicates that osteopontin synthesis can be blocked by inactivation of Akt gene and that metastasis of liver cancer might be inhibited by this intervention.

Objective To explore the mechanisms of TGF-? 1?TGF-? 1RⅡ and NF-?B on the angiogenesis in patients with hepatocellular carcinoma.Methods The expression of TGF-? 1 ?TGF-? RⅡ and NF-?B protein in 36 cases of HCC and surrounding HCC tissue was separately detected using immunohistochemistry technique.To observe the relationship of TGF-? 1 protein and MVD, TGF-? 1RⅡ protein and MVD, NF-?B and TGF-? 1 protein, using CD34 labelling vessel endothelial cell. Results The expression of TGF-? 1 and MVD in HCC tissue was higher than that in surrounding HCC tissue (P

Objective To investigate the correlation between H.pylori strain possessing cagA gene and the formation of gastric lymphoid follicles(GLF),and the effect of H.pylori infection on the occurrence of GLF.Methods Antral biopsy specimens from 655 patients (chronic gastritis:n=479,peptic ulcer:n=176)were used for detection of H.pylori infection and histological analysis. CagA gene was examined in 70 clinical isolates by means of PCR－ amplification. Results The incidence of lymphoid follicles in gastric mucosa is significantly higher(60.14％ ) in patients with H.pylori infection than those without infection(17.06％ ).GLF is easier to be detected in patients with active gastritis than in those with inactive gastritis. There is no significant difference in the presence of GLF among H.pylori associated gastroduodenal diseases,such as chronic gastritis ,gastric ulcer. Moreover, there is also no significant relationship between H.pylori strains possessing cagA gene and GLF. Conclusion The presence of GLF might directly related with H.pylori infection, and can be observed as a constant morpholgical marker of H.pylori related gastroduodenal disease ;The formation of gastric lymphoid follicles is not related to the cytotoxin of cag A gene of H.pylori.

Objectives:To explore a new way in treatment of severe acute pancreatitis(SAP). Methods:From 1995 to 2001,23 patients with SAP proved by clinic and CT were treated, and compared the local artery infusion with intravenous drip on effect,mortality and time of hospitalization. Results:The mortality and time of hospitalization in 12 artery infusion and 11 intravenous drip were (14.4?3.1),(29.3?6.1) days of hospitalization and 8.33%,27.27%(mortality),respectively. Conclusions:The mortality and the time of hospitalization can be reduced by local artery infusion of medicine.

BACKGROUND:Genetic factors, environment, chronic infection, and autoimmune disorders are considered to be involved in the pathogenesis of ankylosing spondylitis. Genetic factors play an important role in the pathogenesis of the disease. Ethnic and regional diversity of differentialy expressed genes has become research hotspot because of family aggregation and ethnic diversity of ankylosing spondylitis.
OBJECTIVE:To screen differentialy expressed genes in Xinjiang Uygur and Han patients with ankylosing spondylitis by microarray screening and compare differences in gene expressions.
METHODS: Uygur and Han patients with active ankylosing spondylitis in department of rheumatology of our hospital were randomly colected with five patients for each. In addition, three healthy volunteers were selected as controls. RNA from peripheral blood was extracted and used for microarray hybridization after probe preparation to screen differentialy expressed genes in ankylosing spondylitis samples and the microarray results were verified by semi-quantitative RT-PCR analysis.
RESULTS AND CONCLUSION: Twenty differentialy expressed miRNAs were screened in Uygur and Han patients with active ankylosing spondylitis (P < 0.05). From relationship analysis of target genes and miRNAs, 15 target genes corresponding to the 79 miRNAs involved in human leucocyte antigens and interleukins which linked to human immunity system were found. These findings suggest that differentialy expressed genes can be screened from Uygur and Han patients with ankylosing spondylitis.

It is difficult for medical students to understand electrocardiogram theory. The sec-ond clinical medical college of Wuhan University has explored clinical pathophysiology and therapy (CPPT) pattern in electrocardiogram education. Basic medical knowledge and clinic medical knowl-edge are combined with electrocardiogram theory to reinforce students ' comprehension and attract theit interest in order to obtain better teaching effect. Drawing themselves, analyzing electrocardiogram sys-tematically and memorizing theory with figure is aimed at solving forgetful problems. In addition, the problems such as lack of conformable teaching material, professional teaching teams and objective mode of examination are raised, and the solutions are explored under CPPT pattern.

Objective To investigate the protective effects and mechanisms of granulocyte-colony-stimulating factor (G-CSF)on a rabbit model of chronic myocardial ischemia.Methods Myocardial ischemia models were created by partial ligation of the left anterior descending coronary artery in Japanese white male rabbits.Rabbits were subcutaneously injected with G-CSF (G-CSF group)or saline (control group)for 6 days after myocardial ischemia.The percentage of CD34-positive cells in the peripheral blood was evaluated by flow cytometry,and CD34-positive cells homing and vWF expression in the ischemic myocardium were determined by immunohistochemistry.Results Rabbits in G-CSF group had a higher survival rate than those in control group (P <0.05).Immunohistochemistry of the ischemic myocardium showed that compared with control group,G-CSF group had increased homing of CD34-positive cells on day 7 post-surgery,and more vessels on day 28 post-surgery by anti-von Willebrand factor staining.In addition,we observed an increase in the percentage of CD34-positive cells in the peripheral blood in G-CSF group.Conclusion G-CSF produces an obvious protective effect against chronic myocardial ischemia in rabbits by increasing stem cell mobilization,homing to ischemic myocardium and accelerating neovascularization.

Objective:To establish an animal model of slow transit constipation and the pathobiological changes in interstitial cell of Cajal in colon. Methods:The mouse model was established by subcutaneous administration of morphine. Fecal weight was recorded daily. Transit functions of intestinal movement were examined by activated charcoal suspension pushing test and the changes of interstitial cell of Cajal were observed by immunohistochemical methods. Results:Compared with the controlled mice, there was a significant decrease in fecal weight daily(P

Objective To investigate the role of different culture densities of embryoid bodies (EBs) in cardiac dif-ferentiation of mouse embryonic stem cells (ESCs). Methods The mouse ESCs were cultured in hanging drops for 3 days, followed by another 2 days for suspension culture to form EBs. Suspended EBs of different densities (60 or 120 EBs/60 mm tissue culture dish) were transferred onto tissue culture plates. The cardiac specific troponin T (TnT) was detected by immu-nocytochemistry. The percentage of beating EBs was calculated at different time points. The mRNA expression of cardiac spe-cific transcriptional factors Nkx2.5, GATA4 and cardiac specific proteinsβ-MHC and ANF were detected by RT-PCR. The protein expressions of TnT and p38 were detected by Western-blot assay. Results Spontaneously beating EBs were posi-tively stained with TnT. There were significantly higher percentage of beating EBs, higher gene expression levels of Nkx 2.5, GATA4,β-MHC and ANF and higher protein expression of TnT in high density culture group than those of low density cul-ture group (P＜0.05). Furthermore, there was significantly higher activity of p38 pathway in high density culture group than that of low density culture group. And the percentages of beating EBs and TnT protein expression were decreased by p 38 pathway inhibitor SB203580. Conclusion The culture density of EBs is important in regulating the cardiac differentiation of ESCs. The high cell-density density culture of EBs enhances the cardiac differentiation of ESCs, which might be mediated by p38 signaling pathway.

This study was aimed to establish an HPLC method to determine three ginsenosides in Shengmai ultra-micro powder. The kromasil C18 (250 mm í 4.6 mm, 5 μm) was used as analytical column. The mobile phase was composed of acetonitrile (A) and water (B) with gradient elution (0~35 min, 19% A; 35~55 min, 19%~29% A; 55~70 min, 29% A; 70~100 min, 29%~40% A) at a flow rate of 1 mL·min-1. The detection wavelength was 203 nm and the column temperature was 30℃. The injection volume was 10 μL. The results showed that the linear ranges of ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1 were 0.083~0.834 μg, 0.086~0.863 μg, 0.091~0.911 μg, respec-tively. The average recovery rates (n = 6) were 100.7%, 100.5%, 100.5%, respectively. It was concluded that this method was quick, sensitive, repeatable and suitable to determine contents of ginsenoside Rg1, ginsenoside Re and ginsenoside Rb1 in Shengmai ultra-micro powder.