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Objective To observe the clinical efficacy of moving cupping in the anterior cervical region plus bloodletting in treating acute pharyngitis.Method Thirty-six patients with acute pharyngitis were randomized into two groups, 18 cases each. The treatment group was intervened by moving cupping in the anterior cervical region plus bloodletting at Tiantu (CV22), and the control group was intervened by oral administration of Pu Di Lanoral liquid. The therapeutic efficacy was evaluated by using the acute pharyngitis symptoms grading scale, and the adverse reactions in the treatment group were also observed.Result The two treatment methods were both effective, and the between-group difference in comparing the total effective rate was statistically insignificant (P>0.05). Compared with the control group, the treatment group showed a swifter action; pharyngeal pain, dry and burning sensations in pharynx, and mucus congestion were effectively improved, the disease duration was shortened, and there were no adverse reactions in the treatment group.Conclusion Moving cupping in the anterior cervical region plus bloodletting can effectively mitigate the sufferings of the patients with acute pharyngitis without producing any adverse reactions.
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Objective@#To conduct an epidemiological investigation of two leptospirosis death cases reported in Guizhou Province in 2014.@*Methods@#The information of the patients were investigated and analyzed. The serological detection, samples of the two patients was detected using ELISA and microscopic agglutination test (MAT). Leptospira carrier status of murine host animal in the living environment of the two patients was investigated in October and November of 2014. Leptospires in the kidney were cultured and isolated, the isolates were identified using Leptospira specific PCR and further identified with serogroup specific PCR and the conventional MAT. The relativity between the carrier status of murine and the death cases of human leptospirosis was analyzed.@*Results@#The two death cases of human leptospirosis came from Liping County and the clinical symptoms were consistent with the diagnosis criteria for Leptospirosis. The results of ELISA detection showed that the anti-Leptospira antibody was positive for one of the death cases, MAT identified the serum reacted with sera-group icterohaemorrhagiae Leptospira, while the serum sample of the other case was failed to perform antibody detection due to hemolysis. 1 600 traps were placed in the living environment of the two death cases and 183 murine rodents were trapped. The murine density was 11.44% (183/1 600); 40 leptospirea suspected strains were isolated and all of them were isolated from Apodemus agrarius. The positive rate was 21.86% (40/183); 95 Apodemus agrarius were trapped and the murine density was 5.93% (95/1 600). Species specific PCR identified all the 40 strains as Leptospire. Serogroup specific PCR further identification showed that they were iterohaemorrahgiae serogroup Leptospria. interrogans.@*Conclusion@#Anti-iterohaemorrahgiae Leptospira antibody was detected from one of the two patients. 40 strains of iterohaemorrahgiae serogroup Leptospira interrogans were isolated and all of them were isolated from Apodemus agrarius in the living environment and the serogroup of the Leptospira matched with the serological detection results from patients, which indicated that the two death cases were caused by the infection of iterohaemorrahgiae serogroup Leptospira interrogans, and Apodemus agrarius were the potential source of infection.
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<p><b>OBJECTIVE</b>To identify and characterize the Brucella strains from Guizhou province in 2010-2013.</p><p><b>METHODS</b>A total of 12 strains of Brucella suspicious bacteria were isolated in Guizhou province from 2010 to 2013. Four strains (GZLL3, GZLL4, GZLL11 and SH2) were isolated from goat blood samples and eight strains (SH4, GZZY, GZSQ, GZZA, BR13001, BR13004, BR13005 and BR13006) were isolated from blood samples of patient 12 Brucella suspicious strains were identified and characterized using conventional methods. Brucella genus specific gene BCSP31-based PCR (BCSP31-PCR) was used to identify the genus of Brucella and IS711 insert sequence-based PCR (AMOS-PCR) was applied to identify the species of Brucella strains. Goats and patients originated Brucella strains were comparatively analysed using Pulse-field Gel Electrophoresis (PFGE).</p><p><b>RESULTS</b>Both of conventional methods and PCR identified the 12 Brucella suspicious strains as B. melitensis biotype 3. BCSP31-PCR identification results showed that a specific DNA bands (223 bp) were detected in all the 12 strains and positive control samples with no DNA band in negative samples. AMOS-PCR amplified a 731 bp-DNA bands in all the 12 strains, with 731 bp, 498 bp and 275 bp in M5, S2 and A19 strains, respectively, and no DNA band was detected in the negative control samples. PFGE analysis showed that 12 Brucella isolates from patients and goats showed consistent PFGE patterns with the digestion of restriction enzyme Xba I.</p><p><b>CONCLUSION</b>The epidemic species/type of Brucella in both human and animal in Guizhou province was B. melitensis biotype 3 and goat was the main animal source of infection of brucellosis in Guizhou province.</p>
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Animals , Humans , Bacterial Typing Techniques , Brucella , Classification , Brucellosis , Epidemiology , China , Epidemiology , DNA, Bacterial , Goats , Molecular Typing , Polymerase Chain ReactionABSTRACT
Objective To evaluate the inhibitory effects of shRNAs targeting genes encoding viral capsomeres VP1-VP4 on enterovirus 71 (EV71) replication when used alone or in combination.Methods Short hairpin RNAs (shRNAs) targeting genes encoding VP1-VP4 protein of EV71 were designed and then respectively inserted into lentiviral vector pLKD-CMV-GFP-U6 to construct the recombinant plasmids.The expression plasmids together with psPAX2 and pMD2.G were transfected into 293T cells to induce the expression of recombinant lentiviruses,which were collected on the third day after transfection.The titers of recombined lentiviruses were determined by real-time PCR.The effects of shRNAs used alone or in combination on the expression of EV71 at mRNA and protein levels were respectively detected by real-time PCR and Western blot.Results The inhibitory effects of shRNAs on EV71 replication showed no significant differences among various strains (isolated from fatal cases,severe cases,mild cases and FY0805) (P＞0.05).Their inhibition rates were 51.6％ (sh-VP1-1),85.1％ (sh-VP1-2),76.4％ (sh-VP1-3),57.5％ (sh-VP2-1),81.4％ (sh-VP2-2),79.5％ (sh-VP2-3),68.9％ (sh-VP3) and 56.7％ (sh-VP4) respectively,and they were in a dosage dependent manner.sh-VP1-2 in combination with sh-VP2-2 showed the highest inhibition rate reaching up to 96.6％.Moreover,shRNAs used in combination showed better effects than any one used alone even at double dosage.Conclusion All shRNAs targeting viral capsid VP1-VP4 genes showed inhibitory effects on EV71 replication with inhibition rates over 50％ and the effects could be strengthened when using shRNAs in combination.
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<p><b>OBJECTIVE</b>To understand the genetic and epidemiologic characteristic of Brucella (B.) melitensis strains isolated in Guizhou province in 2010-2012.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>
Subject(s)
Humans , Brucella melitensis , Genetics , Brucellosis , Epidemiology , China , Epidemiology , Cluster Analysis , Genetic Variation , Minisatellite Repeats , Polymerase Chain ReactionABSTRACT
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.
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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.
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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.
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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.
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Objective To confirm the death of a child injured by a dog was due to rabies and to understand the molecular biologic features of rabies virus in Kaili,Guizhou province.Methods Brain tissue samples of patient and dog were collected to detect the rabies virus by direct immunofluorescence assay (DFA) and RT-nested PCR assay.Homology and phylogenetic tree were analyzed based on the whole nucleotide and deduced amino acid sequence of N gene of rabies virus followed by molecular epidemiological analysis.Results Both the human and dog brain tissue samples were confirmed positive by DFA and RT-nested PCR assay.The homology analysis of N gene sequences among GZH,GZD and other epidemic and vaccine rabies strains isolated from other provinces and other countries indicated that the detected samples shared the highest homology with the strain detected in Anlong prefecture in Guizhou in the year of 2006,and the homology between GZH and GZD was as high as 100％.Besides,among the vaccine strains,GZH and GZD showed the highest homology with strain CNT.Phylogenetic analysis indicated that the two samples were very close and belonged to genetype 1 lyssavirus,with the closest relationship between samples reported in Guizhou and Beijing.Conclusion It was confirmed on the viral molecular level that both the human and dog in Kaili were suffered from rabies,and the pathogens were genetype 1 lyssavirus.The prevalent strains in Kaili city was probably imported from other prefectures of Guizhou province,suggesting that prevention and control measures on rabies in Guizhou province should be strengthened.