AIM:To study the action of nucleoside diphosphate kinase A(NDPK-A)on the growth of S_ 180,H_ 22,Lewis and H460.METHODS:S_ 180 or H_ 22 cell(5?106)were inoculate subcutaneously into the right armpit of 85 Kunming mice,which were randomized into 8 groups.Lewis lung carcinoma cells(2?105)were inoculate subcutaneously into the right armpit of 85 C57BL/6 mice,which were randomized as Kunming mice.From the 2nd day,the treated groups were given different dose of rhNDPK-A once a day for 8 days(for S_ 180 or H_ 22 by iv)or for 10 days(for Lewis by ip),and the control group was given physiological saline only.H460 tissue pieces about 1.5 mm?1.5 mm?1.5 mm each were inoculated subcutaneously into the armpit of 38 Balb/c/neu mice.After the volume of xenograft become 100 mm?100 mm?100 mm,the nude mice were randomized into 5 groups and given different dose of rhNDPK-A once a day for 17 days.2 days after above treatments,the mice were killed and dissected.The knubs were peeled off and weighted.RESULTS:The growth of S_ 180,H_ 22 and H460 were inhibited by rhNDPK and the growth of H_ 22 was inhibited by rhNDPK at dose of 20 mg/kg combined with cisplatin(0.5 mg/kg).But the growth of Lewis lung cancer was not inhibited.CONCLUSION:rhNDPK-A inhibited the growth of S_ 180,H_ 22 and H460.rhNDPK-A(20 mg/kg)potentiated the antitumor action of cisplatin on H_ 22.

AIM: To study the inhibitory effects of nm23-H1 gene on proliferation and invasion of human lung adenocarcinoma A549 cell line. METHODS: Recombinant eukaryotic expression vector pcDNA3.1-nm23-H1 containing full length of human nm23-H1 cDNA was constructed and transfected into a human lung adenocarcinoma A549 cell line by lipofectamine. Cell strain that expressed nm23-H1 stably was screened out by G418 and named pcDNA-nm23-A549. Expression of nm23-H1 was identified by RT-PCR and immunohistochemistry. Growth curves were drawn to detect the inhibitory effects on cell proliferation. Cell cycle of pcDNA-nm23-A549 was examined by flow cytometry. Atomic force microscopy was used to observe the filopodia on the surface of the cells. RESULTS: Introduction of nm23-H1 obviously inhibited the proliferation of A549. Expression of nm23-H1 did not induce apotosis in A549 cells but increased the percentage of phase G_1 cells and decreased phase S cells. Meanwhile, phase G_1 to phase S transition was restrained. Filopodia in the cell surface was much fewer and its structure changed in cells transformed. CONCLUSION: nm23-H1 is capable of inhibiting A549 proliferation and decreasing its metastatic ability, probably by interfering with cell cycle and cell surface structure.

AIM: To construct E. coli expression plasmid of recombinant human NDPK-A with a 6?His tag, optimize the expression condition and identify the activity of the product. METHODS: nm23-H1 was subcloned from plasmid pBVNMH1 to pQE40 which contain 6?His purification tag. The expression condition was modulated in grades to get the optimal expression. We purified protein with the Ni+-NTA affinity chromatography column, identified the immunogenicity of the product with Western blot, and measured the kinases activity with HPLC. In addition, angiogenesis inhibition activity of rhNDPK was identified by CAM. RESULTS: The sequence of nm23-H1 subclone in pQE40 was exactly correct. The expression rate of rhNDPK-A was 49 6%. Purified rhNDPK-A specially recognized the antiserum of NDPK-A. It also inhibited angiogenesis. CONCLUSION: PQE-nm23H1 containing 6?His can express target protein at high level. This purification method is simple than other methods, and the product has the same activity as natural human NDPK-A.

Traditional EP analysis is developed under the condition that the background noises in EP are Gaussian distributed. Alpha stable distribution, a generalization of Gaussian, is better for modeling impulsive noises than Gaussian distribution in biomedical signal processing. Conventional blind separation and estimation method of evoked potentials is based on second order statistics (SOS). In this paper, we propose a new algorithm based on minimum dispersion criterion and Givens matrix. The simulation experiments show that the proposed new algorithm is more robust than the conventional algorithm.
Algorithms ; Artifacts ; Brain ; physiology ; Electroencephalography ; methods ; Evoked Potentials ; physiology ; Humans ; Normal Distribution ; Signal Processing, Computer-Assisted

AIM: To study the pathological change in mouse organs immunitied by inactivated SARS-CoV vaccine. METHODS: Inactivated SARS-CoV vaccine was injected into BALB/c and C57BL/6 mice. Anti-SARS antibody was analyzed by ELISA. After 8 weeks, the immunitied mice were killed and those organs were analyzed by pathological methods. RESULTS: Anti-SARS antibody in mice was positive after 8 days. Only minimal injury was observed in a few lungs and livers, but the other organs were not. CONCLUSIONS: Inactivated SARS-CoV vaccine induced mice to create antibody, whereas they did not cause severe injury. This result will be valuable for vaccine into clinical research. [

Objective To understand the epidemic trend of Tibetan sheep plague in Guoluo Prefecture,Qinghai Province,we detected the plague F1 antibody in Tibetan sheep serum in this area.Methods Indirect hemagglutination test (IHA) and colloidal gold immunochromatography (GICA) were applied to test serum samples of Tibetan sheep which were separated from 5 ml whole blood drew from jugular vein in Maqin County,Maduo County,Gande County,Banma County,Jiuzhi County and Dari County in 2014 and 2015.Results We collected 1 481 serum samples,566 from Maqin County,315 from Maduo County,150 from Gande County,150 from Banma County,150 from Jiuzhi County and 150 from Dari County.Totally 14 serum samples showed F1 antibody positive,the positive rate was 0.95％ (14/1 481),and they were all from Maqin County.Conclusions This area has the prevalence of Tibetan sheep plague.Therefore,the monitoring work of Tibetan sheep plague should be strengthened.

Objective In order to acquaint with the prevalence of Tibetan sheep plague in this area, we conducted a serum epidemiological investigation of Tibetan sheep plague in Qinghai Province. Methods Indirect hemagglutination assay (IHA) and colloidal gold immunochromatography (GICA) were applied to test serum samples of Tibetan sheep and whole blood samples from jugular vein of Tibetan sheep were collected in 8 Prefectures of Qinghai Province from 2013 to 2016. Results A total of 86 positive Tibetan sheep serum samples with plague F1 antibody were detected by both methods, and the positive rate was 0.68％ (86/12710), the samples collected in Xinghai County Hainan Prefecture had the highest positive rate, which was 5.20％ (27/519). The Haixi Prefecture and Yushu Prefecture were historical epidemic areas, the positive rates were 0.65％(15/2313) and 0.26％(6/2293), respectively. Hainan Prefecture, Guoluo Prefacture and Huangnan Prefecture were newly confirmed epidemic areas, the positive rates were 1.61％ (28/1741), 1.01％ (15/1481), and 1.44％(19/1316), respectively. The antibody titers were 1:20 to 1:5120, the samples collected in Maqin County Guoluo Prefecture had the highest titer, namely 1 :5120. Conclusions In Qinghai Province, Tibetan sheep plague is endemic, and there are outbreaks in some regions. So we have to enhance the Tibetan sheep plague monitoring especially in Marmot plague epidemic area.

OBJECTIVETo study the biological and genetic characteristics of 119 strains of Yersinia (Y.) pestis isolated from plague patients in Qinghai province, from 1958-2012.

METHODSBoth regular methods and different region(DFR)molecular typing techniques were used to study the epidemiological characteristics on 119 strains of Y. pesticin Qinghai during 1958-2012. Sources of Y. pestis from two outbreaks, in Nangqian county in 2004 and in Xinghai county in 2009,Qinghai province were also analyzed.

RESULTS105 strains of Y. pestis were identified as Qinghai-Tibet Plateau Ecotype while the other 6 strains as Qilian Mountains Ecotype. 84.03% (100/119) of the tested strains carried 4 virulence factors F1(+), Pst I(+), VW(+) and Pgm(+)). 97.30% (72/74) of the tested strains showed high virulence. Strains that carrying 52×10(6), 65×10(6), 92×10(6) plasmids were distributed in Hainan, Haibei, Haixi,Yushu,Guoluo, Huangnan and Huangyuan counties. Genomovar 5 and 8 were the main gene types that circling around Qinghai Lake. Genomovar 10 was found in strains of Y. pesticin Nangqian county while Genomovar 8 was found in the strains isolated from human plague patient during the epidemics in Xinghai county in Qinghai.

CONCLUSIONData from biological and genetic analyses on the epidemics of human plague in Nangqian county in 2004 and in Xinghai county in 2009 demonstrated that methods as DFR genotyping and virulence factors profiles, as well as plasmids profiles were powerful tools in confirming the human plague epidemics and sources of infection.

China ; epidemiology ; Genotype ; Humans ; Plague ; epidemiology ; microbiology ; Yersinia pestis ; genetics ; isolation & purification

OBJECTIVETo analyze the plasmid features and geographical distribution characteristics of Yersinia pestis of different plague foci in China.

METHODSA total of 2 213 Yersinia pestis strains were colected from 11 Chinese plague foci separated during 1943 to 2012, and plasmid DNA according to alkali cracking method, and measured the relative molecular mass (Mr) of plasmid DNA based on the standard plasmid contrast method, then analyzed the plasmid profiles by agar gel electrophoresis.

RESULTSA total of 2 213 strains had 16 kinds of plasmids with different Mr, including 4×10(6), 6×10(6), 7×10(6), 13×10(6), 16×10(6), 20×10(6), 22×10(6), 23×10(6), 27×10(6), 30×10(6), 36×10(6), 45×10(6), 52×10(6), 65×10(6), 72×10(6) and 90×10(6). Plasmid were classified into 26 kinds of plasmid profiles. A total of 2 213 Yersinia pestis strains contained 4 large plasmids, 52×10(6), 65×10(6), 72×10(6) and 90×10(6), whose ratio was 22.10% (589/2 213), 75.60% (1 672/2 213), 0.17% (4/2 213), 2.12% (47/2 213), respectively. Among which, strains with plasmid 52×10(6), 65×10(6), 90×10(6) distributed in Qinghai-Tibet plateau Himalayan Marmot natural plague foci, strains with 72×10(6) plasmid only distributed in Inner Mongolia Meriones unguiculatus natural plague foci and Junggar Basin R. opimus natural plague foci, and 65×10(6) plasmid distributed in all the other foci.

CONCLUSIONStrains in Chinese 11 plague foci contained 4 kinds of large plasmid, the Mr respectively were 52×10(6), 65×10(6), 72×10(6), 90×10(6), which were classified into 26 kinds of plasmid profiles with other plasmid. These plasmid profiles distributed in relatively independent epidemic focus.

Animals ; China ; Genotype ; Plague ; Plasmids ; Yersinia pestis

OBJECTIVETo identify the epidemiology and etiology characteristics of Tibetan sheep plague in Qinghai plateau.

METHODSThe background materials of Qinghai Tibetan sheep plague found during 1975 to 2009 were summarized, the regional, time and interpersonal distribution, infection routes, ecological factors for the spread were used to analyze; followed by choosing 14 Yersinia pestis strains isolated from such sheep for biochemical test, toxicity test, virulence factors identification, plasmid analysis, and DFR genotype.

RESULTSFrom 1975 to 2009, 14 Yersinia pestis strains were isolated from Tibetan sheep in Qinghai province. Tibetan sheep, as the infection source, had caused 10 cases of human plague, 25 plague patients, and 13 cases of death. All of the initial cases were infected due to eating Tibetan sheep died of plague; followed by cases due to contact of plague patients, while all the initial cases were bubonic plague. Cases of bubonic plague developed into secondary pneumonic plague and septicemia plague were most popular and with high mortality. Most of the Tibetan sheep plague and human plague occurred in Gannan ecological zone in southern Gansu province, which was closely related to its unique ecological and geographical landscape. Tibetan sheep plague coincided with human plague caused by Tibetan sheep, especially noteworthy was that November (a time for marmots to start their dormancy) witnesses the number of Yersinia pestis strains isolated from Tibetan sheep and human plague cases caused by Tibetan sheep. This constituted the underlying cause that the epidemic time of Tibetan sheep plague lags obviously behind that of the Marmot plague. It was confirmed in the study that all the 14 strains were of Qinghai-Tibet Plateau ecotype, with virulence factors evaluation and toxicity test demonstrating strains as velogenic. As found in the (Different Region) DFR genotyping, the strains isolated from Yushu county and Zhiduo county were genomovar 5, the two strain isolated from Nangqian county were genomovar 5 and genomovar 7, while those isolated Delingha region were genomovar 8.

CONCLUSIONTibetan sheep were vulnerable to plague infection, hence causing human plague as the infectious source. The Yersinia pestis strains isolated from Tibetan sheep plague carried pathogen characteristics of Qinghai-Tibet plateau plague, developing many new characteristics of such plague.

Animals ; Ecology ; Genotype ; Geography ; Humans ; Marmota ; Plague ; epidemiology ; veterinary ; Plasmids ; Sheep ; microbiology ; Tibet ; epidemiology ; Yersinia pestis