Bovine aortic endothelial cells (BAECs) were cultured with high glucose (33 mmol/L), 4 mg/L green tea polyphenols (GTPs) or 4 mg/L GTPs co-treatment with high glucose for 24 h in the presence or absence of Bafilomycin-A1 (BAF). We observed that high glucose increased the accumulation of LC3-II. Treatment with BAF did not further increase the accumulation of LC3-II. Results also showed an increased level of p62 and decreased Beclin-1. However, GTPs showed inversed trends of those proteins. Furthermore, GTPs co-treatment with high glucose decreased the level of LC3-II and a much higher accumulation of LC3-II was observed in the presence of BAF in comparison with high glucose alone. Results also showed a decreased p62 and increased Beclin-1. The results demonstrated that GTPs alleviated autophagy inhibition induced by high glucose, which may be involved in the endothelial protective effects of green tea against hyperglycemia.
Animals ; Autophagy ; drug effects ; Cattle ; Cells, Cultured ; Endothelial Cells ; drug effects ; metabolism ; Gene Expression Regulation ; drug effects ; Glucose ; toxicity ; Macrolides ; pharmacology ; Polyphenols ; chemistry ; pharmacology ; Tea ; chemistry

Objective To observe the clinical value of indocyanine green retention rate at 15 minutes (ICGR15) in the evaluation of hepatic functional reserve. Methods A total of 185 patients with liver disease, including 45 cases of liver failure, 90 cases of cirrhosis (child A, B and C, respectively), 20 cases of acute hepatitis, 30 cases of chronic hepatitis (mild, moderate). Expression levels of ICGR15 were compared between groups. Values of ICGR15, total bilirubin (TBIL), albumin (ALB), blood coagulation time (PT) were compared before treatment and one month after treatment in hepatic failure group. Levels of alanine aminotransferase (ALT), aspertate aminotransferase (AST), TBIL and ICGR15 were compared before treatment and 1 month after treatment in acute hepatitis group. Results Levels of ICGR15 (%) were 56.3±14.7, 28.9±9.6, 22.4±6.8 and 13.7±2.3 in liver failure group, liver cirrhosis group, acute hepatitis group and chronic hepatitis group, which showed a gradual downward trend (F=125.317, P<0.05). Among them, the levels of ICGR15 (%) were 17.3±5.4, 25.7±7.5 and 34.5±7.3 in Child A, B and C groups of liver cirrhosis group, which showed a gradual upward trend (P<0.05). After one month treatment, levels of TBIL, PT and ICGR15 were significantly lower than T helper 17 cells; intima-media thickness before the treatment in liver failure group. The levels of ALT, AST, TBIL and ICGR15 were significantly lower after treatment than those before treatment in acute hepatitis group (P<0.05). Conclusion ICGR15 can reflect hepatic reserved function, which is not affected by the application of albumin and fresh plasma, and makes up the deficiency of PT and ALB detection.

OBJECTIVETo understand the genetic and epidemiologic characteristic of Brucella (B.) melitensis strains isolated in Guizhou province in 2010-2012.

METHODSB. genus specific BCSP31-PCR and species-specific AMOS-PCR were used to identify the bacteria strain, while the identified strains were analyzed under MLVA-16 and cluster analysis of B. melitensis strains. The strains were isolated from Guizhou and other provinces.

RESULTSSix B. melitensis strains were identified as B. melitensis using the BCSP31-PCR and AMOS-PCR. Data from the MLVA-16 analysis revealed the differences of repeated numbers at parts of the VNTR locus in the six strains isolated in Guizhou province. The six strains from Guizhou province and 105 B. melitensis strains from other province could be divided into 72 MLVA types(MT). Strain ZY and ZA from Guizhou province were typed as MT63, and LL3, LL4 and LL11 were typed as MT67, while strain SQ was typed as MT72. Data from the clustering analysis showed that ZY,ZA, LL3, LL4 and LL9 were most closely clustered with B. melitensis isolates from Yunnan, Fujian and Guangdong provinces, but strain SQ was genetically remote from other strains.

CONCLUSIONPCR methods, combined with MLVA-16, identified the six B. melitensis strains isolated in Guizhou province in 2010-2012 as B. melitensis biovar 3, with the genetic diversity of the strains showed. Six strains were closely related to the B. melitensis strains from Yunnan, Fujian and Guangdong provinces. The results of this study provided scientific basis for the control and prevention of Brucellosis in Guizhou province.

Brucella melitensis ; genetics ; isolation & purification ; Brucellosis ; epidemiology ; China ; epidemiology ; Cluster Analysis ; Genetic Variation ; Humans ; Minisatellite Repeats ; Polymerase Chain Reaction

Eight patients with suspected cases of C .jejuni were etiologically diagnosed and analyzed in this study to pro-vide scientific basis for the confirmation of the cases of human campylobacteriosis in Guizhou Province ,China .Blood or feces of 8 suspected patients were employed to isolate bacteria strains .Conventional and multi-PCR techniques were applied to identify suspicious bacteria strains .The C .jejuni strains were analyzed by using Pulsed Field Gel Electrophoresis (PFGE) .Suspicious strains of C .jejuni were isolated from all the 8 suspected patients of campylobacteriosis and anticipated genes fragment were detected with multi-PCR .With the digestion of restriction enzyme SmaI ,the 8 C .jejuni strains were divided into 7 PFGE pat-terns with 7-10 DNA bands .Cluster analysis showed that the gross similarity of 8 strains of C . jejuni was more than 50% . The similarity of PFGE patterns between strain GZ201004 and GZ201005 from diarrhea patients was as high as 100% ,while the similarity of strain GZ201201 and GZ201007 was 66 .7% .Moreover ,C . jejuni were detected from all the suspected pa-tients of campylobacteriosis .PFGE results indicated that strains GZ201004 and GZ201005 were from the same source ,while all the 8 isolates showed PFGE polymorphism .

To identify the isolated suspicious strain of Campylobacter jejuni from the blood of bacteremia patient in Guizhou Province ,China ,conventional and molecular techniques (specific mPCR and NAP-mPCR) were used to identify suspi-cious bacteria strains .Results showed that Campylobacter jejuni suspicious colonies were cultured in bacteremia patient blood samples .The strain was identified as Campylobacter jejuni ssp . jejuni by conventional tests and was identified as Campy-lobacter jejuni by genus specific mPCR .Then the strain was classified as Campylobacter jejuni ssp . jejuni by subspecies NAP-mPCR .The strain was identified as Campylobacter jejuni ssp .jejuni isolated from the blood of bacteremia patient and Campylobacter jejuni can be identified subspecies by NAP-mPCR .

Objective To understand the genetic and epidemiologic characteristic of Brucella (B.) melitensis strains isolated in Guizhou province in 2010-2012. Methods B. genus specific BCSP31-PCR and species-specific AMOS-PCR were used to identify the bacteria strain,while the identified strains were analyzed under MLVA-16 and cluster analysis of B. melitensis strains. The strains were isolated from Guizhou and other provinces. Results Six B. melitensis strains were identified as B. melitensis using the BCSP31-PCR and AMOS-PCR. Data from the MLVA-16 analysis revealed the differences of repeated numbers at parts of the VNTR locus in the six strains isolated in Guizhou province. The six strains from Guizhou province and 105 B. melitensis strains from other province could be divided into 72 MLVA types(MT). Strain ZY and ZA from Guizhou province were typed as MT63,and LL3,LL4 and LL11 were typed as MT67,while strain SQ was typed as MT72. Data from the clustering analysis showed that ZY,ZA,LL3,LL4 and LL9 were most closely clustered with B. melitensis isolates from Yunnan,Fujian and Guangdong provinces,but strain SQ was genetically remote from other strains. Conclusion PCR methods,combined with MLVA-16, identified the six B. melitensis strains isolated in Guizhou province in 2010-2012 as B. melitensis biovar 3,with the genetic diversity of the strains showed. Six strains were closely related to the B. melitensis strains from Yunnan,Fujian and Guangdong provinces. The results of this study provided scientific basis for the control and prevention of Brucellosis in Guizhou province.

To study the characteristic of glycoprotein gene sequence of rabies virus in Guizhou Province in recent years and provide scientific basis for effective control and prevention of rabies,RT-PCR assay were used to detect human and dog brain tissues derived from different prefectures of Guizhou.The amplification products were sequenced and analyzed with bioinformatics software.The results showed that the full-length G gene sequences of 8 positive samples were obtained by RT-PCR amplification,sequencing and splicing.The homology of eight G gene sequences from Guizhou Province were between 87.4％ -99.9％ and 83.3.％- 100％ on nucleotide and deduced amino acid level,respectively,and the highest homology were found with the genotype 1 strains ( 86.5 ％- 87.0％ for nucleotide and 83.3 ％- 100 ％ for amino acid) among genotype 1- 7 representative strains.Besides,phylogenetic analysis based on the G gene indicated that the relationship of 8 strains derived from Guizhou were closest to genotype 1 Lyssavirus,and the strains of Guizhou were very close to the strains come from Hubei,Hunan,Anhui,Guangxi,Jiangsu provinces and Shanghai,except for GZ01 and GZ09.Moreover,GZ09 were evolutionarily closed to the strains come from Malaysia and Thailand,while the remaining sequences were closed to the strain of Indonesia.This study confirmed on the G gene level that rabies virus epidemic strains circulated in Guizhou Province in recent years belonged to rabies virus genotype 1 and the evolutionary relationship with reported strains come from different provinces of China and different countries were revealed in this study.It will provide scientific basis for effective control and prevention of human and animal rabies in Guizhou Province.

Objective To study the differences of glycoprotein gene (G gene) between rabies virus epidemic strains of Guizhou province in recent years and vaccine strains,and to provide scientific basis for the development of rabies vaccine and establishment of effective control and prevention measures.Methods RT-PCR assay was used to amplify the G gene of rabies positive brain tissues samples of human and dog derived from Guizhou province in recent years.The amplification products were sequenced and comparatively analyzed with that of vaccine strains by using bioinformatics software.Results Eight full-length G gene sequences were obtained by RT-PCR amplification,sequencing and splicing.The homogeny of G gene between 8 epidemic strains of Guizhou province and 9 vaccine strains were 82.0％-94.1％ and 87.6％-97.5％ on nucleotide and deduced amino acid level,respectively,and the highest homogeny were found with the human vaccine strain CTN (87.0％-94.1％ for nucleotide and 93.7.％-97.5％ for amino acid) among the 6 human rabies vaccine strains,while highest homogeny were found with strain Flury (83.9％-84.6％ for nucleotide and 91.1％-93.0％ for amino acid) among the three animal vaccine strains.Besides,among the 8 epidemic strains from Guizhou province,strain GZ09 collected in the year of 2005 was of the highest homogeny with human rabies vaccine strain CTN and animal rabies vaccine strain Flury,while strain GZ30 collected in the year of 2010 was of lowest homogeny with human rabies vaccine strain CTN and animal rabies vaccine strain Flury.Moreover,phylogenetic analysis based on the G gene indicated that the relationship of 8 epidemic strains derived from Guizhou,the 9 vaccine strains and genotype 1 Lyssavirus were clustered to a same branch.Vaccine strain CTN among the 9 vaccine strains was closest to the 8 epidemic strains,and the other 8 vaccine strains were relatively more distant from the epidemic strains of Guizhou province.In addition,phylogenetic analysis indicated that among the 8 epidemic strains from Guizhou province,strain GZ09 collectcd in the year of 2005 was of closest evolutionary relationship to CTN,while the other 7 epidemic strains were relatively more distant from CTN.Conclusion This study confirmed on the G gene level that rabies virus strains circulated in Guizhou province in recent years and the vaccine strains used in China belonged to rabies virus genotype 1,and the virus strains circulated in Guizhou province in recent years is of smallest difference with the human vaccine strain CTN and animal vaccine strain Flury.Besides,as time goes on,the difference between the epidemic strain and the vaccine strains becomes more and more obvious.The results of this study will provide scientific basis for the development of rabies vaccine and establishment of effective control and prevention measures.

Objective To study the composition of variant and point mutations of Norovirus GⅡ.4 in Guiyang regions.Methods From June to November 2010,cases information and fecal specimens were collected from guard-hospitals in Guiyang regions,who had caught the acute-gastroenteritis.Noroviruses in specimens were detected by a real-time reverse transcription polymerase chain reaction(real-time RT-PCR),and then partial genotyped norovirus-positive clinical samples (in random) were cloned and sequenced in VP1 gene code.Furthermore,the gene sequences were compared with the published variants at home and abroad of norovirus(GⅡ.4),including the phylogenetic analyses of genomes and variation of amino acids within individual sites.Results Those 267 specimens were GⅡ-norovirus-positive(62.68％) in 426 clinical samples.There were nine GⅡ.4-norovirus-positive VP1 gene-sequences available,and two subtype-norovirus variants (GⅡ.4 2008a and G Ⅱ.4 2008b variant) were epidemic in 2010,Guizhou province.The homology between and in subgroups were 95.90％-96.72％ and 99.45％-100％.Two amino acids within individual sites were apt to mutate.Conclusion Norovirus GⅡ genotype were predominant in summer and fall acute gastroenteritis in 2010 for Guiyang regions,and the variants were diversity.