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Objective@#To investigate the interaction of Ca2+ protein TRPC1 and STIM1 in extracellular Ca2+ -sensing receptor (CaR)-induced extracellular Ca2+ influx and the production of nitric oxide (NO).@*Methods@#Human umbilical vein endothelial cells (HUVECs) were cultured and incubated with CaR agonist spermine (activating store-operates cation channels (SOC) and receptor-operated channels (ROC)), CaR negative allosteric modulator Calhex231 (blocking SOC, activating ROC) and ROC analogue TPA (activating ROC, blocking SOC), protein kinase C (PKC) inhibitor Ro31-8220, PKCs and PKCμ inhibitor Go6967(activate SOC, blocking ROC), respectively. The interaction of TRPC1 and STIM1 was determined using the immunofluorescence methods. The interaction between TRPC1 and STIM1 were examined by Co-immuno precipitation. The HUVECs were divided into: TRPC1 and STIM1 short hairpin RNA group (shTRPC1+ shSTIM1 group), vehicle-TRPC1+ vehicle-STIM1 group and control group. The cells were incubated with four different treatments under the action of above mentioned interventions, intracellular Ca2+ concentration ([Ca2+ ]i) was detected using the fluorescence Ca2+ indicator Fura-2/AM, the production of NO was determined by DAF-FM.@*Results@#(1) The expression of TRPC1 and STIM1 proteins levels in HUVECs: Under the confocal microscope, TRPC1 and STIM1 protein expression showed masculine gender, both located in cytoplasm in the normal control group. Post incubation with Calhex231+ TPA, Ro31-8220 and Go6967, TRPC1 and STIM1 positioned in cytoplasm was significantly reduced, and the combined TRPC1 and STIM1 was also significantly reduced. (2) The interaction of TRPC1 and STIM1 in HUVECs: The relative ratios of Calhex231+ TPA+ Spermine+ Ca2+ group, Ro31-8220+ Spermine+ Ca2+ group and Go6976+ Spermine+ Ca2+ group STIM1/TRPC1 and TRPC1/STIM1 were as follows: (25.98±2.17)% and (44.10±4.01)%, (20.85±1.01)% and (46.31±3.47)%, (23.88±2.05)% and (39.65±2.91)%, which were significantly lower than those in the control group (100.00±4.66)% and (100.00±6.40)% and in the Spermine+ Ca2+ group (106.04±2.45)% and (107.78±2.66)% (all P<0.05). (3) The influence of joint TRPC1 and STIM1 transfection to four different drugs treated HUVECs on [Ca2+ ]i and NO generation: The changes of two excitation fluorescence intensity ratio and NO net fluorescence intensity values were consistent, [Ca2+ ]i and NO net fluorescence intensity values were significantly lower in the experimental group than the control group and the vehicle group (all P<0.05), while which were similar between the vehicle group and control group (all P>0.05).@*Conclusions@#Our results indicate that TRPC1 and STIM1 jointly regulate CaR-mediated Ca2+ influx and nitric oxide generation in HUVECs in the form of binary complex.
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Objective@#To investigate the prevalence and characteristics of pathogenic variants in complement genes in Han Chinese children with atypical hemolytic uremic syndrome (aHUS).@*Method@#Eleven Han Chinese children with aHUS, including 9 boys and 2 girls aged between 1 year and 4 months and 13 years, were investigated in Department of Pediatrics, Fuzhou General Hospital, from November 1998 to February 2014. Analysis of variants of all the exons of 10 complement genes (CFH, MCP, CFI, C3, CFB, CFHR1, CFHR2, CFHR3, CFHR4 and CFHR5), including 25 bases from 3′ end and 25 bases from 5' end, was performed in the 11 cases by targeted sequence capture and next generation sequencing. Significant variants detected by next generation sequencing were confirmed by Sanger sequencing. To understand pathogenicity of variants found in the captured genes, we investigated genetic conservation by multiple protein sequence alignment among different species, and analyzed whether the variants were located in protein domains or not, and investigated functional significance by functional computational prediction methods.@*Result@#Twenty-seven percent of Han Chinese children with aHUS carried pathogenic variants in the 10 complement genes. Pathogenic variant CFB 221G>A (R74H) was detected in Patient 3 and Patient 9, which was not found in parents of Patient 3′ , and was found in healthy father of patient 9. Pathogenic variant CFHR5 242C>T (P81L) was found in Patient 2, and was found in healthy father of patient 2. However, no pathogenic variants in genes CFH, MCP, CFI, C3, CFHR1, CFHR2, CFHR3 and CFHR4 were identified.@*Conclusion@#Pathogenic variants in the 10 complement genes were identified in 3/11 of Han Chinese children with aHUS in our study and CFB was the most frequently mutated gene.
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The possibility of heterologous expression of human Stearoyl-CoA Desaturase (scd1) was investigated. The scd1 encoding sequence was inserted into the pNZ8149 to generate the pNZ8149-scd1 expression plasmids. Then we introduced the pNZ8149-scd1 construct into the Lactococcus lactis NZ3900 to investigate its enzyme activity. The results show that heterologous expressed SCD1 enzyme resulted in a 92%-169% increase in the C16:1n-7 and a 53-127% increase in the C18:1n-7 (P<0.05). The SCD1 enzyme was capable of producing n-7 fatty acids in Lactococcus lactis efficiently. It also suggests that the fatty acid desaturases can be heterologous expressed in Lactococcus lactis to produce the helpful fatty acids.
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Humans , Electroporation , Lactococcus lactis , Genetics , Metabolism , Mutagenesis, Insertional , Nisin , Pharmacology , Recombinant Proteins , Genetics , Stearoyl-CoA Desaturase , GeneticsABSTRACT
Objective To explore the epidemical pattern of acute schistosomiasis in Poyang Lake area so as to provide the scientific evidence for control-strategy-making. Methods The data of acute schistosomiasis in 12 counties along Poyang Lake from 1960 to 2007, including the data of 2 special investigations from 1982 to 1984 and from 1987 to 1989 were collected and analyzed with the retrospective investigation methods. Results There were 29 864 cases of acute schistosomiasis in 12 counties of Poyang Lake area from 1960 to 2007, accounting for 85.5% of the total acute schistosomiasis cases of Jiangxi Province. The largest number of cases existed in Yugan County, accounting for 18. 7%. The peak of acute schistosomiasis prevalence was in 1987 with 2 088 cases. However, the case of acute schistosomiasis declined significantly, keeping at a low level, since 1990. During the 2 special investigations, the people whose age were lower than 30 years old, were the susceptible populations of acute schistosomiasis (86. 1% and 82. 9% , respectively) , most of them were peasants and students, and the infection rates were 57. 4% and 21.2%, 61.7% and 21. 8%, respectively. Fishing and swimming were the main infection ways, with the infection rates of 43.0% and 44.9% , respectively. The peak season of infection was from April to October. Conclusions The river bank and continent of Poyang Lake are the main infection areas, the spring and autumn are the main epidemic seasons and the susceptible populations are the peasants, fishmen and young students. It is very important to enhance the comprehensive prevention and treatment measures in the key areas and population for schistosomiasis control.
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Objective To summarize the achievement and experience in leprosy control in Jiangxi Province and provide scientific basis for formulation of leprosy strategies. Methods Based on the National Leprosy Recording and Reporting System, epidemiological data of leprosy in Jiangxi Province were analyzed using computer. Results Jiangxi Province was a leprosy medium endemic area with a higher distribution of leprosy in the south than the north. The prevalence, incidence and detection rates were highest in the 1959s or 1960s and had decreased annually since then. Average incidence rate for the recent 5 years (0.1604 per 100 000) and prevalence rate (0.049 per 10 000) in 1998 reached the criteria of basic elimination of leprosy at provincial level, however, still 15 counties or cities not reaching the criteria. Conclusion The comprehensive measures including early detection of cases, immediate treatment with multidrug therapy and effective health education are helpful and important for leprosy control.
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As a result of observation in lab. ,it showed that the rice straw attracted snail well,it at-tracted uninfected and infected snails, and adult and young snails respectively without singnificant difference. On this basis,we can investigate the snails under water by rice straw curtains during water raising season. This paper reported that the method for weaving rice straw curtain fixed by stone,and using foamed plastics as buoy were easy and convenient. The rate of snail-attraction was highest when it took 14 hours ,especially in August.