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.

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.

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 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.

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

AIM: To analyze the subcellular localization of Arnt2 in rat cerebellar granule neurons (CGNs). METHODS: Based on the amino acids sequence of Arnt2 (LOCUS:NP_036913), the subcellular localization of Arnt2 in eukaryotic cells and the nuclear export signals (NES) of Arnt2 were predicted in CBS bioinformatics database. The subcellular localization of Arnt2 in rat cerebellar granule neurons was detected by the method of laser scanning confocal microscopy (LSM) analysis. RESULTS: It was predicted that Arnt2 located in nuclei of eukaryotic cells with the most probability, while located in cytoplasmic mitochondria with a slight possibility. A nuclear export signal was found in Arnt2 amino acids sequence, it was identified to be the leucine of No.143 that located in N-terminal of Arnt2 amino acids sequence. Finally, the result of LSM analysis shows nuclear localization of Arnt2 in rat CGNs. CONCLUSION: Arnt2 is located in nuclei of normal rat CGNs, it suggests that Arnt2 has the tendency to translocate into mitochondria after induced by some of inducible factors, for both the possibility of mitochondria localization and NES exist in Arnt2 amino acids sequence.

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 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.

AIM: To confirm the differential expression genes in the rat ischemic brain. METHODS: The middle cerebral artery occlusion ischemic model was set up in rats. Fluorescence differential display reverse transcriptase-polymerase chain reaction (FDD RT-PCR) and reverse Northern blotting were used to fast confirm the differential expression genes. RESULTS: Nine differential expression sequence tags, including 6 known sequences and 3 unknown sequences, were confirmed. Among the known sequences, mus musculus ab1-interactor1,homo sapiens CGI-99 protein, tissue inhibitor of metalloproteinase 3 and homosapiens nuclear receptor co-repressor were up-regulated while homo-sapiens nuclear matrix protein p84 and coatomer protein complex, subunit gamma 2 were down-regulated. CONCLUSIONS: ① Combination of fluorescence differential display reverse transcriptase-polymerase chain reaction (FDD RT-PCR) and reverse Northern blotting is a method to fast-confirm the differential expression genes; ② There are differential expression genes in ischemic brain regions compared to non-ischemic parts. [

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.