It is known that no restriction endonuclease cleavage site exists at 12 codon in N-ras gene and at 12 and 13 codon in K-ras gene.Amismatched base was incorporated at the 3' end primer to creat a kind of restriction endonuclease cleavage site.Then it was possible for us to analyze the point mutation of the above mentioned codons with the polymerase chain reaction-restriction fragment length polymorphism.In this paper,the study of the point mutation at 12 and 61 codon in c-Ha-ras,at 12 codon in N-ras,and at 12 and 13 codon in K-ras was reported.

BACKGROUND: Increasing evidence identifies the immune system not as an isolated system with automodulations, but one that interacts with the central nervous system.OBJECTIVE: To investigate the distribution of c-fos expression in the lung and brain tissues of asthmatic rats and explore is significance.DESIGN: Randomized controlled study.SETTING: Department of Oncology, Southern Hospital, and Department of Neurosurgery, Zhujiang Hospital, Southern Medical University.MATERIALS: The experiment was conducted Department of Oncology,Southern Hospital, and Department of Neurosurgery, Zhujiang Hospital,Southern Medical University, between January and August 2004. Fourteen healthy male rats were randomized into experimental group (n=10) and control group (n=4).METHODS: On the first day of experiment, the rats in experimental group received intraperitoneal injection of 2 mL of the suspension containing 10 mg albumen, 200 mg aluminum hydroxide powder and inactivated pertussis vaccine (5×109), and subjected to inhalation of ultrasonically atomized 10 g/L albumen from on the 15th day, 2 times per hour for totally 3 days, to induce asthma in the rats. The rats in the control group received intraperitoneal injection of 2 mL normal saline on the 1st day and inhalation of ultrasonically normal saline on the 15th day, 30 mL a day for totally 3 days. The lung and brain tissues of all the anesthetized rats were fixed by perfusion, and immunohistochemical method with ovin-biotin-peroxidase complex and imaging analysis system were used to observe the distribution of Fos protein in the lung and brain.MAIN OUTCOME MEASURES: Distribution of c-Fos protein in lung and cerebrum.c-Fos in the lung and brain tissues was obviously higher in asthmatic group than in the control group (P ＜ 0.05), located mainly in the parietal-fontal cortex, limbic forebrain (cingulum cortex, pyriform cortex and central amygdaloid nucleus and so on), thalamus paraventricular nucleus, hypothalamus paraventricular nucleus, supraoptic nucleus, lateral region of the hypothalamus, hypothalamus periventricular nucleus, nucleus of solitary tract,area postrema and ventrolateral medulla. No obvious Fos expression was observed in the cerebellum. A large number of c-Fos-positive cells were observed in the wall of the airway and lungs in asthmatic rats, mainly distributing in the mucous membrane and submucous layer and around the smooth muscles; in the control rats, no positive cells or only occasional cells with weak c-Fos positivity were found in the wall of the airway and lungs.CONCLUSION: c-Fos expression increases obviously not only in the lungs of asthmatic rats, but also in medullar and its ascending projecting nuclei (hypothalamus, amygdala and so on), suggesting that the expression of protooncogene c-fos might be closely related with neuroimmunomodulation in asthma.

Immuno-blot was used in diagnosis of schistosomiasis japonica using diagnostic 31/32 KD proteins in S.japonicum Among 52 confirmed schistosemiasis cases 100% showed positive reaction while none of the sera of 20 normal control reacted. No cross reactions occured with sera from 36 paragonimiasis; 26 cysticercosis and 50 trichinelliasis cases. However, 11.5-58.3%cross reactions were found with above sera in IHA and ELISA. The diagnostic value of immuno-blot and 31/32 kD schistosome proteins were confirmed.A field application of immuno-blot in diagnosis of schistosomiasis japonica was performed. The positive rates in immuno-blot and IHA were 58.1% and 54.4% respectively in 136 cohorts with and without past history of schistosomiasis in the endemic area. It was found that immuno-blot was highly sensitive, specific and reproducible.It was easy to perform and suitable for field use.

Objective To understand the moving patterns of Oncomelania snails, intermediate host of S.japonicum, in the water bodies. . Methods. Based on the biological features of the snails, methods and techniques in relation to hydraulics and silt engineering were adopted to investigate the active scrawl ability and passive movement of the snails. . Results and Conclusion . The scrawl speed of the snails themselves was very low, 2.^45 mm per minute only. The active movement in the water current was therefore almost negligible. The major moving patterns of the snails in rivers and canals were that: 1. Snails adsorbed on different kinds of carriers drifted on the water. 2. Snails suspended on the water and drifted with the current, these were the young snails under 7 weeks of age. Adult snail showed a strong ability of adhesion, 12 times higher than its body weight.

AIM: To observe the effect of E2F decoy DNA on proliferation and apoptosis of androgen-independent prostate cancer cell line PC-3M.METHODS: E2F decoy DNA,ARE decoy DNA and control decoy DNA were transfected into PC-3M cells with lipofectamine,respectively.Their effects on cell proliferation were detected by MTT assay.The changes of cell morphology were observed by inverted phase contrast microscope.The cell apoptotic rate was determined by flow cytometry(FCM) analysis and chromosome DNA ladder was detected by DNA gel electrophoresis.The expression of c-Myc mRNA and cyclin D1 mRNA was detected by RT-PCR.The protein levels of c-Myc and cyclin D1 were detected by Western blotting.RESULTS: The growth of PC-3M cells was inhibited after transfection.The transfected PC-3M cells displayed typical apoptotic morphological changes.The apoptotic rate was 26.35% and DNA ladder was observed after transfection.The expression of c-Myc and cyclin D1 were inhibited.CONCLUSION: These results indicate that E2F decoy DNA induces apoptosis of androgen-independent prostate cancer cell lines PC-3M and inhibits cell proliferation via inhibiting expression of c-Myc and cyclin D1.

Objective To detect the expressions of angiotensin-receptor-1 (AT1R) and hypoxia-inducible factor (HIF)-1αin glomeruli and juxtaglomerular apparatus of different types of lupus nephritis (LN) patients, and analyze the correlation between them with systemic lupus erythematosus disease activity index (SLEDAI) complement 3, serum creatinine and 24-hour proteinuria in order to explore the role of the two factors in the pathogenesis of lupus nephritis (LN). Methods Between May 2010 and April 2016, a total of 90 patients with LN and 8 healthy controls were selected from Department of Rheumatology, Qujing Affiliated Hospital of Kunming Medical University and the First Affiliated Hospital of Kunming Medical University. The expressions of AT1R and HIF-1αin renal biopsy specimens were measured by streptavidin-perosidase (SP) of immunohistochemical stains. Pathological graphic analysis system was used for semi-quantitative estimate. Levels of SLEDAI, C3, serum creatinin and 24-hour proteinuria were also detected. Finally the relationshipbetween the two factors with clinical data was analyzed. The ANOVA test was used for intergroup comparison, and SNK-q test was used for the two groups comparison. Pearson's analysis was used for correlation analysis. Results The AT1R [(10.55 ±0.31)% vs (7.04 ±0.11)%] and HIF-1α [(10.51 ±0.52)% vs (8.96 ±0.31)%] in the glomeruli of typeⅠLN was significantly higher than healthy controls(all P<0.05). In the early phase of LN, RAS was activated and tissues were ischemic and hypoxic. The highest expression of AT1R (18.22 ± 2.11)% and HIF-1α (19.48 ±0.61)% in glomeruli was found in type Ⅳ LN, especially in juxtaglomerular apparatus, AT1R (19.98 ±0.21)% and HIF-1α(24.90 ±0.70)%. AT1R was positively correlated with HIF-1αin the glomer-ulus (r=0.949, P<0.01) and juxtaglomerular apparatus (r=0.762, P<0.05). AT1R and HIF-1αin juxtaglomerular apparatus was positively correlated with 24-hour proteinuria (r=0.756, P<0.05 and r=0.802, P<0.05). Conclusion High expressions of AT1R and HIF-1α have been shown in active LN biopsies. It proves that RAS is activated by ischemia and hypoxia, then it up-regulates HIF-1α expression. Our results suggest that the two factors may be associated with disease activity of LN.