Objective To investigate the effect of lentivirus-mediated siRNA interference of USP39 on proliferation and migration of mice vascular smooth muscle cell in vitro.Methods Five siRNAs of siControl, siRNAUSP39-70, siRNAUSP39-71, siRNAUSP39-72 and siRNAUSP39-73 were designed and sythezied,mice VSMCs were infected with the lentivirus for delivering siRNAUSP39-73, and the stably transfected cells were selected by puromycin.The interference efficiency of siRNAUSP39-73 was assessed with Western blot.The effect of USP39 interference on the proliferation of VSMCs was determined by cells counting and MTT assay.Transwell assay was used to detect the migration of VSMCs.Results Recombinant lentiviral vector siRNAUSP39-73 was successfully transfected into mice VSMCs.Comparing with siControl group and Normal group, USP39 protein level of siRNAUSP39-73 VSMCs were decreased(P<0.05), and the proliferation and migration ability were all inhibited(P<0.05).Conclusion Targeted down-regulation of USP39 expression can inhibit the proliferation and migration of mice VSMCs in vitro.

The growth and development of plant is affected by environmental factors that include light,temperature and water. Many cis-elements that are response to these elicitors interact with transcription factors, and regulate the expression of target genes. It concludes by pointing out recent studies of cis-elements and transcription factors in the plant environmental response promoters, such as light, temperature and water inducible promoter. The molecular mechanisms of gene expression regulated by environmental factors was discussed. This is important to study the mechanisms that plant adapt the environment.

Objective To evaluate the effect of comprehensive control for schistosomiasis with emphasis on environmental modification in the Bianmin River water system of Nanjing City.so as to provide scientific evidence for making up further control measures in this water system.Methods Schistosome infections of Oncomelania snails in the waterway.sentinel mice in water and neighbouring human were investigated longitudinally from 1998 to 2007,and the changes of huaman infection rates in differentyears,the infection rates of sentinel mice and snails in different settings were analyzed and compared.Results A total of 77 395 snails collected from the Bianmin River water system were dissected from 1998 to 2007,and among them,27 snails were infected with Schistosoma japonicum,with a total snail infection rate of 0.03%.A total of 61 039 snails collected from the neighbouring marshland which connected to the Yangtze River wore dissected,and among them,257 were infected with S.japonicum,with a total snail infection rate of 0.42%,and there was a significant difference compared with that in the water system(χ~2=248.55,P<0.01).After the protection works in the waterway,the infection rates of sentinel mice in the water system decreased from 69.68% in 1998 to 17.50% in 2001.with a reduction rate of 74.89%.Two years afterthe clearance ofmarshlandinthewaterway,no infected sentinel mouse was found.The infection rates of residents from 1998 to 2007 were 1.96%,1.37%,1.34%,1.60%,0.30%, 0.26%,0.16%,0.10%,0.04% and 0,respectively,andthe rates declined year by year afterthecomprehensive control.Conclusions The control measures based on the elimination of snail habitats in the waterway that is connected to the Yangtze River have achieved obvious effect.However,the clearance of the re-emerging snail habitats should be carried out termly to consolidate the control effect.

Actins ; metabolism ; Adrenal Cortex Neoplasms ; metabolism ; pathology ; surgery ; Adrenocortical Adenoma ; metabolism ; pathology ; surgery ; Antigens, CD34 ; metabolism ; Carcinoma, Renal Cell ; metabolism ; pathology ; surgery ; Follow-Up Studies ; Hemangioma ; metabolism ; pathology ; surgery ; Humans ; Kidney Neoplasms ; metabolism ; pathology ; surgery ; Male ; Middle Aged ; Neoplasms, Multiple Primary ; metabolism ; pathology ; surgery ; Nephrectomy ; Pelvic Neoplasms ; metabolism ; pathology ; surgery

Adolescent ; Adult ; Alprostadil ; pharmacology ; Anticoagulants ; pharmacology ; Child ; Child, Preschool ; Dextrans ; pharmacology ; Female ; Fibrinolytic Agents ; pharmacology ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Heparin ; pharmacology ; Hepatic Veno-Occlusive Disease ; etiology ; prevention & control ; therapy ; Humans ; Infant ; Male ; Middle Aged ; Platelet Aggregation Inhibitors ; pharmacology ; Treatment Outcome

AIM: To Screen and identify differentially expressed genes that involved in apoptosis model in rat cerebellar granule neurons (CGNs).METHODS: The rat cerebellar granule neurons were isolated and primarily cultured. Fluorescent differential display RT-PCR (FDD RT-PCR) was performed to screen differentially expressed ESTs in the apoptosis model of primarily cultured rat CGNs. ESTs were subcloned into pGEM-T EasyTM vector and then sequenced. Alignment assay in non-redunant database was applied for encoding information. Reverse Northern blotting was used to appraise the results from DDRT-PCR.RESULTS: 164 pieces of differentially expressed ESTs were obtained by FDDRT-PCR. 17 of them were subcloned and sequenced. 5 ESTs of 17 were confirmed to be positive results by reverse Northern blotting. CONCLUSION: DD-PCR is a rapid, simple-operation and sensitive method for screening differentially expressed genes, which would contribute to the molecular mechanisms of apoptosis/survive of CGNs.

AIM: To observe the expression of neuronal Aryl hydrocarbon receptor nuclear translocator 2 (ARNT2) involved in neuronal apoptosis evoked by low K + and to investigate the relationship between ARNT2 and neuronal apoptosis. METHODS: After neuron apoptosis model was established, the changes of mRNA and protein of ARNT2 during apoptosis were investigated by RT-PCR and Western blotting, respectively. Immunofluorescence was analyzed by confocal microscopy to probe the subcellular localization of ARNT2. RESULTS: Induced by low K +, the expression of ARNT2 mRNA was up-regulated obviously at the point of 30 min, and peaks at the point of 1 h. This up-regulated expression lasted for 12 h, and the variation of ARNT2 protein was similar to that of mRNA. The results of immunofluorescence analyzed by confocal microscopy showed that the localization of ARNT2 protein was in the nucleus. CONCLUSION: ARNT2 locate in nuclei of normal cerebellar granule neurons of rat. During the process of apoptosis evoked by low K +, both mRNA and protein of ARNT2 are overexpressed.

AIM To construct a non-normalized cDNA library from Agkistrodon acutus venom gland as an imtial step to develop new and more effective venom by genetic engineering technique for screening and expressing target genes. METHODS The total RNA was extracted from fresh venom gland using Trizol. mRNA was reversely transcripted to cDNA using superscriptⅡ reverse transcriptase. Second-strand synthesis was performed using DNA polymeraseⅠ. After adding EcoRⅠ adaptor, phosphorylating the end and digesting with XhoⅠ, the cDNA was collected in five fractions (<0.25 kb, 0.25-0.5 kb, 0.5-1 kb, 1-2 kb and >2 kb) using the QIAquick Gel Extraction kit and ligated to pBluescriptⅡ vectors. The five libraries obtained were plated by infecting E.coli DH10B, constructing a cDNA library of Agkistrodon acutus venom gland. Sequencing clones at random, 8696 high quality 5′ end expressed sequenced tags (ESTs) were obtained and analyzed. The initial sequences were assembled into 2855 clusters. Among which, one of the clusters (Agkihagin) consisting of 74 ESTs was identified as a novel metalloprtoteinase based on RT-PCR and sequence analysis. RESULTSThe titers of library were 2.048×106. The novel metalloproteinase belonged to PⅢ type metalloproteinase. Its open reading frame was composed of 1827 nucleotides and coded a pre-zymogen of 608 amino acid with zinc-binding domain for metalloproteinase and Asp-Glu-Cys-Asp(DECD) domain for disintegrin. CONCLUSION The capacity of cDNA library of venom gland is above the general level of cDNA library. It would be a helpful platform to construct a catalog for transcripts in the venom gland of the Agkistrodon acutus. The sequence analysis indicates that the deduced amino acid sequence of the identified gene for metalloproteinase share the highest 87% identity with the metalloproteinase genes of other snakes in the GenBank. It lays a good foundation for the study of structure-function relationships of snake venom metalloproteinases.

Objective To explore the roles of macrophage inflammatory protein-1?(MIP-1?)in hypoxic-ischemic encephalopathy(HIE)of newborn infarnts.Methods Serum samples were obtained in 24,72 h and 7 d after birth respectively from 34 newborn infants with HIE,and 20 newborn infants without HIE as control group.Enzyme-linked immunosorbent assay(ELISA)method was used to determine the serum concentrations of MIP-1?.Results Levels of MIP-1? in newborn infants with HIE [(12.47?2.51)ng/L]were significantly higher than that of newborn infants without HIE [(8.63?2.63)ng/L](P0.05).Conclusions MIP-1? are involved in HIE of neonates,and the more severe damage,the higher levels in serum,which suggests that,as an inflammatory mediator,the MIP-1? may play an important role in involvement of brain hypoxic-ischemic damage.

AIM: To clone the full-length of 75A EST. METHODS: After the extraction of total RNA from primary cultured rat cerebellar granule cell of 7DIV in the medium containing 25 mmol/L KCl, T_4 DNA ligase-mediated 5' RACE was used to retrieve 5' unknown sequence of 75A EST, and the first round 5' RACE PCR product was subcloned into pGEM-T easy vector for sequence and homogeneous analysis. RESULTS: The first round of 5' RACE produce a 2.5 kb band, and 75A EST was identified to be partial sequence of Neuron-derived orphan receptor (Nor1) gene. After two more rounds RACE, we firstly cloned the full-length of Nor-1 cDNA. CONCLUSION: T_4 DNA ligase mediated 5' RACE is an efficient method to retrieve information about the 5' termini of mRNAs, and lay a foundation for further study which role Nor1 play in the cerebellar granule cell differentiation or survive.