Objective To study the distribution status and clinical significance of human papillomavirus(HPV) infection geno‐types in condyloma acuminate(CA) tissues of vulva ,vagina and cervix .Methods The gene‐chips combined with polymerase chain reaction (PCR) technology were utilized for detecting 23 kinds of HPV genotypes in tissue specimens from 63 cases of vulval CA , 61 cases of vaginal CA and 65 cases of cervical CA .Their clinical pathological data were analyzed .Results In 63 cases of vulval CA ,56 cases were HPV positive with the HPV infection rate of 88 .89% (56/63) ,in 61 cases of vaginal CA ,55 cases were HPV positive with the HPV infection rate of 90 .16% (55/61) ,and in 65 cases of cervical CA ,62 cases were HPV positive with the HPV infection rate of 95 .39% (62/65) .Conclusion HPV infection is closely related to the CA pathgenesis in vulva ,vagina and cervix . HPV6 and HPV 11 are main stream genotypes ,in which vulval CA is most common .The gene‐chips combined with PCR technology is a method suitable for HPV typing diagnosis ,and has the characteristics of good sensitivity and high specificity ,which has an im‐portant significance for clinical diagnosis ,treatment and vaccine study of CA in femal vulva ,vagina and cervix .

Objective To compare the sensitivity of fluorescent quantitation polymerase chain reaction (fluorescent quantitation method) and gene‐chips typing method(gene‐chips method) in the detection of human papillomavirus(HPV) ,and to analyse differ‐ences and clinical significance .Methods A total of 246 women were selected as subjects ,among them ,111 cases of cervical exfolia‐ted cells and 135 cases of cervical tissues were collected and detected .15 kinds of high‐risk HPV genetypes were detected in all sub‐jects by using fluorescent quantitation method and gene‐chips method respectively ,and the detection results were compared . Results The sensitivity of the fluorescent quantitation method in detecting HPV was 55 .28% and that of the gene‐chips method was 55 .69% ,there was no statistically significant difference in sensitivity between the two methods (P>0 .05) .The two methods had relative high conformance(κ=0 .745) .The positive rate of HPV infection was increased with the progression of cervical dis‐ease .Conclusion The fluorescent quantitation method and the gene‐chips method have a relative high conformance ,and both with high sensitivity in detecting HPV .The severity degree of cervical cytological and histological changes may be positively correlated with HPV infection .

To analyze the molecular basis of the variation of the pathogenicity of the influenza B virus, we rescued a recombinant virus with a deletion in the carboxyl terminal of the NS1 protein using reverse genetics based on the parental virus B-S9 of B/Yamagata/16/88. A mutant strain with a deletion of 171 amino acids in the carboxyl terminal of the NS1 protein was named "B-L5". BALB/c mice were inoculated with 3 X 105 EID50 of B-L5 and the parental virus B-S9, respectively. Then, weight changes, survival, and viral titers were documented. During 3 days post-inoculation (dpi) to 7 dpi, the weight of mice infected with B-S9 decreased. However, the weight of mice infected with B-L5 showed weight decreases only at 2 dpi, and quickly recovered at 3 dpi. B-S9 and B-L5 could replicate in the lungs of BALB/c mice. However, viral titers in the lungs of mice infected with B-L5 were 7900-times lower than those of mice infected with B-S9 at 3 dpi. Viral titers in the lungs of mice infected with B-L5 were not detected at 6 dpi. These results showed that, compared with the parent virus B-S9, the mutant virus B-L5 showed lower pathogenicity in BALB/c mice. Our study suggests that deletion of the carboxyl terminal of the NS1 protein decreases the pathogenicity of the influenza B virus. Establishment of a reverse-genetics system for the B influenza virus will provide a platform for studying its pathogenesis, and mechanism of transmission, and for developing live-attenuated influenza B virus vaccines.
Animals ; Body Weight ; Dogs ; Female ; HEK293 Cells ; Humans ; Influenza B virus ; genetics ; pathogenicity ; physiology ; Madin Darby Canine Kidney Cells ; Mice ; Mice, Inbred BALB C ; Sequence Deletion ; Survival Analysis ; Viral Load ; genetics ; Viral Nonstructural Proteins ; chemistry ; genetics ; Virulence

Purpose To compare the distribution of 23 kinds of human papillomavirus ( HPV) genotypes in tissues of condyloma acu-minata ( CA) of cervix in 120 women and its clinical significance. Methods Polymerase chain reaction ( PCR) and gene-chips tech-nology were utilized for the detection of 23 kinds of HPV genotypes in tissue specimens from 120 cases of CA in cervix and related ma-terials of all subjects were conducted and analyzed. Results There were 115 positive cases in 120 women with CA in cervix and the rate of total HPV infection was 95. 83% (115/120). The rate of single type was 70. 83% (85/120) and multiple types was 25. 00%(30/120). The predominant type of single infection was HPV11 and the infective rate was 45. 00% (54/120), followed by HPV6 (22. 50%, 27/120). Otherwise, the predominant type of multiple infections was HPV6+11 with the infective rate of 20. 00% (6/30), and HPV11+16 infection accounted for 10. 00% (3/30). Conclusions HPV11, 6, 6+11 and 11+16 are the main genotypes in the pathogenesis of CA in cervix in 120 women. PCR and gene-chip technology can detect single and multiple HPV genotyping in tis-sues of CA in cervix with high sensitivity and specificity. Detection of HPV genotypes could be used to understand the prevalence situa-tion of HPV infection in tissues of CA and tumors of cervix and further to provide references for the research and development of HPV vaccine in women.

Objective To investigate the distribution situation of human papillomavirus (HPV) genotypes profile in cervical cells among women in Xuzhou area and its clinical significance .Methods 23 kinds of HPV DNA were extracted in cervical cell samples from 8 010 women in Xiuzhou area .The gene‐chips technique of PCR combined with reverse dot blot was adopted to detect the HPV genotypes .Results Among 8010 cervical cell samples ,there were 1 852 HPV infected cases ,the total HPV infection rate was 23 .12% ,the HPV infection rates of single type accounted for 17 .17% and its predominant types were 16 type (4 .35% ) ,followed by 58 type (2 .12% ) and 52 type (1 .82% ) ,The detection rate of multiple HPV infection was 5 .96% ,in which the predominant types were HPV16+58(4 .40% ) ,16+52(2 .94% ) ,11+16(2 .52% ) .Conclusion The single HPV infection of HPV16 ,58 ,52 and the multiple HPV infection of HPV16+58 ,16+52 ,11+16 are the main genotypes of cervical cells among women in Xuzhou area , this gene chip technique is suitable for the cervical cell sample ,its once detection can detect 23 kinds of HPV genotypes with high specificity and high sensitivity ,which has an important significance for the molecular epidemiologic survey study of HPV genotypes distribution among women in our country .

Objective To study the genotypes of human papillomavirus (HPV)infection in female anus and anal canal condylo-ma acuminata(CA)tissues and their clinical significance.Methods 23 kinds of HPV-DNA were extracted from the paraffin-embed-ded anus and anal canal tissue samples in 140 cases of female CA and detected by using PCR combined with the gene-chips tech-nique.Furthermore the related clinical pathological data of the patients were analyzed.Results Among 140 female anus and anal ca-nal CA tissue samples,103 cases were HPV positive and the total HPV infection rate was 73.57%(103/140).Among them,68 ca-ses were single type HPV infection,the positive detection rate was 48.57%(68/140)and 35 cases were multiple types HPV infec-tion,the positive detection rate was 25.00% (35/140).In single type HPV infection,34 cases were HPV11 and the positive detec-tion rate was 24.29% (34/140),HPV11 was the main infection type,followed by HPV 6 in 27 cases,its positive detection rate was 19.29%(27/140).In the multiple types HPV infection,13 cases were HPV 6 + 11,accounting for 37.14% (13/35 )of multiple types infection,followed by HPV11 +18 in 3 cases and HPV 6+11+16 in 3 cases,each accounting for 8.57%(3/35)of the multi-ple types infection.Conclusion HPV 6,11 ,6+11,11 +18 and 6+11+16 are the main infection genotypes in female anus and anal canal CA.PCR combined with the gene-chips technique is a diagnostic method more suitable for clinical development of HPV geno-typing detection,which has high sensitivity and good specificity and is especially suitable for the molecular epidemiology study of HPV infection.

Objective To explore the clinical distribution states of human papillomavirus genotypes in tissues of 691 women with vulva condyloma acuminates in Nanjing city and Zhenjiang city in Jiangsu Province and genotyping clinical significance.Methods Polymerase chain reaction(PCR)and gene-chips technology were utilized for the detection of 23 kinds of HPV genotypes in tissue specimens from 619 women of vulva condyloma acuminates in Nanjing city and Zhenjiang city in Jiangsu Province.And related materials of all subjects were analyzed.Results In 691 women of vulva condyloma acuminates,597 women of HPV infecton,total infection rate of HPV was 86.40％(597/691),including single genotype infection rate of HPV was 51.38％(355/691),11、6 and 16 genotypes are the most common in single genotypes,they are successively 51.55％(183/355)、41.97％(149/355)and 3.38％(12/355).multiple genotypes infection rate of HPV was 35.02％(242/691),6+11、11+18、6+16 and 11+16 genotypes are the most common in multiple genotypes,they are successively 9.92％(24/242)、9.09％(22/242)、4.96％(12/242)and 4.13％(10/242).Conclusion The low-risk HPV types are the main factors to cause the female vulva CA,a few high-risk HPV types may cause warts as well in tissues of women with vulva condyloma acuminates in Nanjing city and Zhenjiang city in Jiangsu Province.The vulva examine of HPV types should be held to the vulva CA patients.This precaution will has extremely important meaning to the prevention and treatment of the female vulva CA and cervical lesion in our nation.

Objective To compare the distribution situation of human papillomavirus(HPV)infective genotypes in normal cells and atypical squamous cells of undetermined significance(ASC-US)in uterine cervix and its clinical significance.Methods The pol-ymerase chain reaction(PCR)combined with the gene-chips technology were adopted to detect 23 kinds of HPV genotype from 1 000 cases of normal cells specimens and 229 cases of ASC-US specimens.Results 106 cases of HPV-positive infection were de-tected from 1 000 cases of normal cells with the total HPV infection rate of 10.60%(106/1 000),in which the single genotype in-fection rate was 9.30%(93/1 000)and the multiple genotypes infection rate was 1.30%(13/1 000);116 cases of HPV-positive in-fection were detected from 229 cases of cervial ASC-US specimens with the total HPV positive rate was 50.66% (116/229 ),in which the single genotype infection rate was 34.06%(78/229)and the multiple genotypes infection rate was 16.59%(38/229).The total HPV positive rates,single and multiple genotype infection had statistically significantly differences between the two groups(P<0.05).Conclusion The HPV types 16,18,33,42,43,52,58 are the predominant genotypes in normal cervical cells and ASC-US. PCR combined with the gene-chip technology can be used in the HPV genotype detect in cervical cells,conduces to perform the fur-ther distribution management on ASC-US and has the important significance to prevention and control of cervical cancer.

Objective To investigate the distribution of 39 kinds of human papillomavirus(HPV)infection genotypes in female cervical cells and its clinical significance.Methods 39 types of HPV DNA were extracted from 434 samples of female cervical cells. The gene amplification combined with the gene chip technique was adopted to detect 39 kinds of HPV genotype.And the clinical da-ta of the patients were analyzed.Results Among 434 samples of female cervical cell,175 cases were HPV positive,the total HPV infection rate was 40.32%(175/434).Among them,105 cases were the single type HPV infection with the positive detection rate of 24.19%(105/434)and 70 cases were the multiple types HPV infection with the positive detection rate of 16.13%(70/434).Among single type HPV infection,31 cases were the HPV18 infection with the positive detection rate of 17.71%(31/175),which was the main HPV infection type;followed by HPV16 in 12 cases with the positive detection rate of 6.86%(12/175)and HPV52 in 11 cases with the positive detection rate of 6.29%(11/175).Among the multi-type HPV infection,each 2 cases were HPV 6+54,HPV 18+52,HPV 51+68 infection respectively,each accounted for 2.86% of the multi-type HPV infection,which were the main infection types.Conclusion HPV 16,18,52 and HPV 6+54,HPV 18 +52 and HPV 51 +68 are the main HPV infection genotypes of fe-male cervical cells.The gene amplification combined with the gene chips technique is a method suitable for clinically conducting the HPV genotyping diagnosis and the molecular epidemiologic research of HPV infection.Along with the increase of detected HPV genotypes,the HPV infection rate is also increased,its genotypes combinations trend towards diversification.

OBJECTIVETo study the cloning and expression of flagellin gene from Chinese Borrelia burgdorferi, PD91 strain and to evaluate the feasibility of using recombinant protein as diagnostic antigen when comparing the gene sequence with flagellin gene from North American Borrelia burgdorferi B31.

METHODSThe piece of genes coding flagellin from Chinese Borrelia burgdorferi PD91 by polymerase chain reaction (PCR) method was obtained, and constructed recombinant plasmid, before transformed into E. coli BL21 strain, and induced. The recombinant plasmid was identified with enzyme cutoff and gene sequence comparison. Efficient expression strain was selected and the expression product was analyzed with sodium amplified polymorphic-polyacrylamide gel electrophoresis (SDS-PAGE) and Western-blot method.

RESULTSThe recombinant protein (r-flagellin) expressed in host bacteria was successful. By means of western-blot assay, the immunological response showed the same antigenicity between r-flagellin and PD91 flagellin. The piece of genes coding flagellin of PD91 was 1011 bp, but when comparing with that of North American Borrelia burgdorferi it showed 94.70% homology. Homology between the sequence of amino acid of the r-flagellin and that of B31 flagellin was 95.85%.

CONCLUSIONFlagellin gene of Borrelia garinii of Chinese Lyme disease spirochete was successfully cloned and expressed for the first time. It was proved that the immunoreactivity of r-flagellin was the same as the natural flagellin.

Amino Acid Sequence ; Base Sequence ; Borrelia burgdorferi ; genetics ; isolation & purification ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Flagellin ; biosynthesis ; genetics ; Humans ; Lyme Disease ; microbiology ; Molecular Sequence Data ; Plasmids ; genetics ; Recombinant Proteins ; biosynthesis ; genetics