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Objective@#To analyze the genetic diversity of Aedes albopictus populations in the coastal areas of southern China by using the microsatellite markers to provide a basis for the control of vectors.@*Methods@#Genetic diversity and clustering analysis of Aedes albopictus populations were studied in the 7 microsatellite loci, in Hangzhou, Ningbo and Yiwu of Zhejiang province, Longyan of Fujian province, Guangzhou of Guangdong province, Nanning of Guangxi Zhuang Autonomous Region and Haikou of Hainan province.@*Results@#Numbers of different alleles (5.429-7.571), effective alleles (2.897-3.632), allele richness (5.236-7.170) and expected heterozygosity (0.538- 0.637) were detected from each of the Aedes albopictus population by using 7 microsatellite markers. The inbreeding coefficients appeared as 0.008-0.332, with heterozygote deficiency, in these populations. Fixation index of the whole populations was 0.058, suggesting that the genetic variation among the 7 populations was 5.8%. Data from the Neighbor-Joining clustering analysis showed that populations from Hangzhou and Yiwu belonged to one branch while Longyan and Guangzhou populations constituted another branch. Aedes albopictus populations of Nanning and Haikou showed great genetic variation but formed a single branch. Bayesian analysis on Aedes albopictus populations showed that the possible number of clusters was 3.@*Conclusions@#Based on 7 microsatellite loci, relatively high genetic diversity and medium level of genetic differentiation that increasing with the geographical distances, were found in these Aedes albopictus populations, from the coastal areas in southern China.
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Objective@# To apply DNA barcoding to identifying the rodents in Zhejiang Province. @*Methods @#Rodents were captured from Jiashan,Longyou,Yunhe and Ninghai counties in Zhejiang Province. The DNA was extracted from ears of rodent samples,and was amplified and sequenced with mitochondrial cytochrome C oxidase subunit I（COI）genes. The obtained sequences were compared with the related sequences in GenBank,and neighbour-joining evolutionary tree was constructed. Then the results by DNA barcoding and by morphological identification were compared. @*Results @#A total of 22 COI gene samples were amplified. The evolutionary tree constructed by 18 samples was consistent with the morphological identification results and 4 samples were different：Suncus murinus should be Crocidura lasiura,infant rats of Rattus losea and Rattus tanezumi was re-identified as Rattus rattus,infant rats of Microtus fortis（sample number：NH-1）needs further identification. @*Conclusion @#DNA barcoding can effectively correct the errors of morphological identification,thus combining the two methods could improve the accuracy of rodent identification.
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Objective To investigate the effects of small interfering RNA (siRNA) silencing Nanog gene on the ability of migration and invasion of the human lung adenocarcinoma A549 cells. Methods The human lung adenocarcinoma A549 cells were transfected with siRNA targeting Nanog gene , and three experiment groups were set up. The expression level of Nanog was detected using reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis. Cell migration was examined by wound healing assay and cell invasion was detected by Transwell assay. Results The Nanog silencing cell group (A549-siNanog) showed much lower level of Nanog mRNA and protein (0.40 ± 0.06, 0.50 ± 0.03) than A549-siNC cell group (0.97 ± 0.03, 0.85 ± 0.02; P < 0.05) under RT-PCR and Western blot analysis. Meanwhile, the wounded area filled rate and the number of invaded cells of A549-siNanog cell group (57% ± 0.04, 69.60 ± 17.14) were decreased significantly compared to A549-siNC cell group (95% ± 0.02, 209.60 ± 15.40; P < 0.05). Conclusion siRNA targeting human Nanog could specially suppress the expression of Nanog gene in lung adenocarcinoma A549 cells. In this way, it couldsignificantly reduce the capability of migration and invasion of A549 cells.
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Objective To study the effect of the inhibitor of apoptosis protein,Livin on proliferation and multi?drug resistance of lung adenocarcino?ma cells A549. Methods A549 cells were transfected with the eukaryotic expression vector pcDNA3.1?Livin. A549 cell clone with stable expres?sion of Livin was obtained through G418 screening. Expressions of Livin mRNA and protein in the transfected cells were respectively measured by re?verse transcription polymerase chain reaction(RT?PCR)and Western blot. The distribution of cell cycle phase was determined using flow cytometry. The level of P?gp mRNA and protein in A549 cells transfected with pcDNA3.1?Livin was detected by RT?PCR and Western blot. The analysis of multi?drug resistance of A549 treated with different chemotherapeutics was performed by MTT. Results The mRNA and protein expressions of Liv?in were both significantly increased in the transfected A549 cells. The flow cytometry analysis showed there was higher percentage of S phase and low?er percentage of G0/G1 phase in A549 cells transfected with pcDNA3.1?Livin. Compared with control groups,the expression of P?gp mRNA and pro?tein was increased in A549 cells transfected with pcDNA3.1?Livin,which showed a higher drug resistance and lower sensitivity to chemotherapic drugs such as ADM,MTX,CTX,and DDP(P<0.05). Conclusion Overexpression of Livin could enhance the proliferation of A549 cells,and high expression of P?gp caused by Livin could serve as one of the causes for multi?drug resistance in lung adenocarcinoma against chemotherapies.
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Objective To observe the multi-segmental cervical spondylotic myelopathy with simple anterior or posterior joint pre-and post-operative prognosis of spinal cord function improved and the status, explore co-operation after the clinical efficacy and complications. Methods The clinical data of 298 cases of multi-segmental cervical spondylotic myelopathy with anterior or posterior of the simple pre-and post-joint surgery from January 2001 to January 2008 were retrospectively analyzed. The clinical efficacy, titanium anterior cervical decompression and fusion surgery net implanted titanium plate fixation 121 cases, posterior open-door laminoplasty 112 cases, 65 cases of combined surgery before and after. JOA score line of spinal cord function and somatosensory evoked potential, as compared 3 groups after surgical efficacy. Results All patients were followed-up 1- 7 years, averaged (4.7±1.4) years. The anterior cervical decompression and fusion surgery titanium mesh implanted titanium plate fixation to improve the rate was 78.1%, excellent and good rate was 72.7%(88/121). Posterior open-door laminoplasty to improve the rate was 70.6%, excellent and good rate was 66.1% (74/112), there was statistically significant between them (P < 0.05). After anterior surgery, improving rate was 86.7%, excellent and good rate was 83.1% (54/65). Anterior and posterior combined surgery before and after comparison was significant (P < 0.05), regardless of near-term results, long-term effects were better than that of anterior or posterior surgery. Conclusions The spinal cord in the treatment of multi-segmental operation of cervical spondylosis after anterior surgery is obviously superior to the efficacy of anterior or posterior surgery alone. Spinal stability anterior, posterior, after a joint operation before the lower one by one.
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The ampC ?-lactamases gene in Escherichia coli(E.coli) is different from other Gram-negative bacteria.E.coli contains a chromosomal ampC gene which has a weak promoter as well as a transcriptional attenuator.The promoter of the ampC gene in E.coli is part of the preceding frd operon,the attenuator of the ampC gene is a transcription terminator for the frd operon.The ampC regulatory gene,ampR,is absent.Strains carrying the wild-type gene produce a low basal amount of AmpC.Studies on the molecular basis of AmpC overproduction in E.coli have shown that some hyperproducers contain mutation in the promoter region and/or attenautor and/or ampC-coding region of ampC,while others contain more than one copy of ampC.Acquisition of a stronger promoter or insertion of an insertion element containing promoter sequences or regulatory gene ampR has also been proposed as the molecular basis of hyperproduction of AmpC in some E.coli strains.Plasmid-mediated AmpC ?-lactamases have been discovered frequently in E.coli strains.This is another reason for hyperproduction of AmpC ?-lactamases.