1.Molecular analysis of two strains of hantavirus isolated from Zhejiang province
Pingping YAO ; Hanping ZHU ; Fang XU ; Zhiyong ZHU
Chinese Journal of Laboratory Medicine 2003;0(07):-
Objective To compare the molecular characteristics of hantanvirus strains isolated from 1980 to 2002 in Zhejiang Province by PCR and nucleotide sequencing.Methods Total RNA was extracted from hantanvirus infected Vero-E6 cells. The M segment cDNA of hantaan ZJ4 and ZJ7 strains were obtained by reverse transcription and polymerase chain reaction, subsequently cloned into pUCm-T vector and sequencing.Results Two strains ZJ4 and ZJ7 of hantanvirus isolated from Rottus in Zhejiang province were HTN viruses. HTN virus strain Z10 was isolated in 1980. Comparison ZJ4 and ZJ7 with Z10 strains indicated that there were 88.3% and 92.3% homology at the nucleotide level.Conclusion The HTN virus were very conserved. The homology of HTN virus isolated from Zhejiang Province was high nucleotide level, though they were isolated at more than 20-year intervals.
2.Expression of matrix metalloproteinase 2 and its correlation with tumor microvascular density and tumor metastasis in nasopharyngeal carcinoma
Li YAO ; Xing LU ; Pingping LIU ; Hongyi HU ; Fengan LIU
Chinese Archives of Otolaryngology-Head and Neck Surgery 2006;0(10):-
OBJECTIVE To study the expression of matrix metalloproteinase 2 (MMP-2) and its correlation with microvascular density(MVD) and tumor metastasis in nasopharyngeal carcinoma. METHODS The expression of MMP-2 and MVD were detected in the specimens of 50 cases with nasopharyngeal carcinoma and 15 cases with nasopharyngeal inflammation by immunohistochemical technique. RESULTS The MVD in nasopharyngeal carcinoma(21.92?7.80) was significantly higher than that in inflammation nasopharyngeal tissues (9.23? 1.84, P
3.Effects and primary mechanism of arctigenin in C6 rat glioma
Qinyong SU ; Xiaomei LI ; Jingchun YAO ; Pingping WANG ; Guimin ZHANG
Chinese Pharmacological Bulletin 2015;(6):805-809
Aim To observe the effect and primary mechanism of arctigenin ( ARG) in C6 rat glioma. At the same time, to investigate the effect of ARG com-bined with temozolomide. Methods C6 glioma rat model was established, and 90 rats were divided into six groups, which were subcutaneously administered with model, low and high ARG (0. 05 and 0. 1 mg· kg-1 , sc) , temozolomide (20 mg·kg-1 , p. o. ) , low ARG combined with temozolomide(TMZ / ARG 0. 05) and high ARG combined with temozolomide ( TMZ /ARG 0. 1 ) . The tumor specimens of brain were col-lected after tumor graft. Proliferating cell nuclear anti-gen ( PCNA ) , glial fibrillary acidic protein ( GFAP ) and CD40 in tumor specimens were determined by im-munohistochemistry. Results ① Compared with the model group, the tumor sizes of rats in the arctigenin treatment groups were decreased ( P<0. 05 ) . ②ARG
significantly decreased PCNA and CD40 expression ( P<0. 05 ) and increased GFAP expression ( P<0. 05 ) .③ Compared with model group, arctigenin combined with temozolomide decreased the tumor sizes ( P <0. 01 ) , and the tumor inhibition rate was higher than that of the arctigenin and temozolomide. At the same time, arctigenin combined with temozolomide de-creased PCNA and CD40 expression ( P <0. 01 ) and increased GFAP expression ( P <0. 05 ) , which was better than arctigenin and temozolomide. Conclusion Arctigenin inhibits rat glioma growth, and synergizes with temozolomide, which may be associated with in-hibiting PCNA and CD40 expression and strengthening GFAP expression.
4.Molecular cloning and nucleotide sequence analysis of the S-segment in Hantavirus isolated in Zhejiang province
Fang XU ; Pingping YAO ; Hanping ZHU ; Ronghui XIE
Chinese Journal of Zoonoses 2009;(7):665-668
One strain of Hantavirus(HV) was isolated from patients with hemorrhagic fever with renal syndrome (HFRS) in Zhejiang province and its S-segment was cloned and submitted to nucleotide sequence analysis in order to determine the type of strain and extent of genetic variation for further study on its evolution and mutation .The HV antigen was detected from mouse lungs in endemic area by direct immunofluorescene test and the HV-positive sample of mouse lung was inoculated to Vero-E6 cells to isolate virus. The total cellular RNA was extracted from infected cell culture and the S segment gene was amplicated by RT-PCR. Then, the purified PCR product was cloned into T vector for sequencing. The result showed that the full-length sequence of the S segment was 1 700 bp with one open reading frame (ORF) encoding 429 amino acids. Comparison with Hantavirus HTN type showed 85.7%-91.9% homology at the nucleotide level. In comparison with the SEO type of viruses, the homology at nucleotide level was shown to be 71.2%-75%. This new HV strain may be typed as to HTN virus and may be a new subtype of this virus.
5.Sequence analyses of the Cps2J gene in 9 strains of Streptococcus suis serotype2 isolated in Zhejiang province
Pingping YAO ; Fusu WANG ; Hanping ZHU ; Lingling MEI ; Juyian YE ; Jin LUO ; Zheng ZHANG ; Chenhui YAO
Chinese Journal of Zoonoses 2010;(1):62-64
To explore the molecular characteristics of Streptococcus suis serotype 2(ss2) isolated in Zhejiang province by deciding the variation loci and its variation frequency of Cps2J gene.The total DNA of 9 strains of ss2 isolated in Zhejiang province were extracted and amplifed by PCR. Then,the Cps2J fragments were cloned into plasmid carrier and completely sequenced after purification.Finally,the sequence results of all 9 ss2 isolates were compared with those obtained by other studies around the world.It was found that the open reading fragments of Cps2J in 9 SS2 isolates encoding 333 amino acids were 999 bp in length.Comparisons of this region among ss2 isolates revealed a similarity of between 98.8% and 99.9%, while the homology to ss1 strains varied between 56.8% and 57.0%.Our study shows the sequences of complete Cps2J segment are fairly stable and all these 9 ss2 strains of different sources possibly have the same evolutionary origin.
6.Full-length nucleotide sequence analysis of the S and M segments in Z5 strain of Hantavirus Z5 strain
Chan LI ; Ronghui XIE ; Hanping ZHU ; Fang XU ; Pingping YAO ; Yinkai CHENG ; Zhiyong ZHU
Chinese Journal of Zoonoses 2010;(3):215-217,225
The M and S segment cDNAs of hantavirus Z5 strain was amplified by RT-PCR,and the purified PCR products were cloned into vector pGEM-T and then sequenced.It was demonstrated that the M genome segment of Z5 was found to be 3 616 nucleotides in length with a single open reading frame encoding 1 135 amino acids.And the S genome segment was 1700 nucleotides in length with a single open reading frame encoding 429 amino acids.As demonstrated by the homologous analysis of nucleotides and amino acids,it was showed that the Z5 strain belonged to hantaan viruses HTN type and was the same subtype of the Z10 strain.It is conclouded that difference in nucleotide sequence exists between Z5 strain with other Hantavirus strains but high level of homology in amino acid sequences is still present.
7.Complete genome sequence analysis of the Hantavirus ZT71 strain
Ronghui XIE ; Jingqing WENG ; Pingping YAO ; Fang XU ; Haiping ZHU ; Zhiyong ZHU
Chinese Journal of Microbiology and Immunology 2008;28(2):149-153
Objective To study the complete genome sequence of Hantavirus ZT71 strain gene isolated in Zhejiang province and explore its evolution. nethods The total RNA was prepared from ZT71 virus infected cells and the RT-PCR products were cloned into T vector, sequenced and analyzed. Results The L, M and S segments of the strain ZT71 genome were 6530,3651 and 1753 nucleotides in length with a single open reading frame individually encoding 2151,1133 and 429 amino acids. The sequence analysis of nucleotides showed that the homology of L, M and S segments of strain ZT71 between those of other strains of Seoul virus could reach 95.5%-99.7%, 84.1%-99.6% and 88.7%-99.5%, respectively. The analysis of the deduced amino acids showed the similar result. The source of strain ZT71 could be traced from the analysis of the phylogentic trees of nucleotides and amino acids, and it should belong to Seoul type of Hantavirus which was also verified serologically. Conclusion The nucleotide sequences and deduced amino acid sequences of L, M and S segments of strain ZT71 are similar to that of those of Seoul type of Hantavirus. And Hantaan type virus used to be prevalent primarily in Zhejiang province,and it would be an endemic area of mixed type of Hantavirus since the discoveries of the viruses of Soeul type in recent years.
8.Expression of tissue inhibitor of metalloproteinase-2 in nasopharyngeal carcinoma.
Li YAO ; Pingping LIU ; Xing LU ; Fengan LIU
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2010;24(18):831-833
OBJECTIVE:
To study the expression of tissue inhibitor of matrix metalloproteinase 2 (TIMP2)in the nasopharyngeal carcinoma and to evaluate the relationships between the expression of TIMP2 and biological behavior of NPC.
METHOD:
The expression of TIMP2 was observed by immunohistochemical technique in 50 cases with nasopharyngeal carcinoma and 15 cases of Inflammation nasopharyngeal tissues.
RESULT:
The positive rates of TIMP2 are significantly higher in NPC than in Inflammation tissues (P < 0.05). The expression of TIMP2 increase significantly with lymph node metastasis (P < 0.01), but had no relevant to the rise clinical invasion stage (P > 0.05).
CONCLUSION
The results indicate that TIMP2 may play a critical role in assisting carcinoma cell to degrade basement membrane to invade surrounding tissues and to form metastatic colonies in lymph node. Especially the unbalance in MMP2/TIMP2 has probably a prognostic value in nasopharyngeal carcinoma.
Adult
;
Double-Blind Method
;
Female
;
Humans
;
Male
;
Matrix Metalloproteinase 2
;
metabolism
;
Middle Aged
;
Nasopharyngeal Neoplasms
;
metabolism
;
pathology
;
Neoplasm Staging
;
Tissue Inhibitor of Metalloproteinase-2
;
metabolism
;
Young Adult
10.The host investigation and virus isolation of hantavirus in Tiantai County
Pingping YAO ; Fang XU ; HanPing ZHU ; Zhangnyu YANG ; Yisheng SUN ; Hangjing LU ; Chen CHEN ; Weilong PANG ; Yun ZHANG ; Haiqing XIANG
Journal of Preventive Medicine 2019;31(5):433-436
Objective:
To learn the population,virus status and viral types of hantavirus(HV)hosts in Tiantai County of Zhejiang Province from 2011 to 2018,and to provide evidence for hemorrhagic fever with renal syndrome(HFRS)control.
Methods:
Rodents in Tiantai County were captured by night trapping method. After the species and age of rodents were identified,the composition of rodent species,dominant species and density of rodents were analyzed. The lungs and blood of rodents were sampled to detect the antigen and antibody of HV by immunofluorescence method. The HV antigen-positive lung samples were detected by RT-PCR with specific primers of HV S fragment,then HV was isolated and identified by inoculating Vero-E6 cells.
Results:
The average rodent density in Tiantai County from 2011 to 2018 was 4.44%. The rodent density in the field and residential areas were 4.94% and 2.23%,respectively. Ten species of rodents were identified,with Apodemus agrarius dominant in the field and Rattus norvegicus in the residential areas. Sixty-seven lung samples were HV antigen positive(4.13%),one from Rattus norvegicus and the other sixty-six from Apodemus agrarius. Seventy-nine blood samples were HV antibody positive(4.86%),all from Apodemus agrarius. Thirty-four HV antigen-positive lung samples were positive(50.75%)after RT-PCR amplification. Twenty-two strains of virus were isolated and all of them were from Apodemus agrarius,including twenty-one strains of Hantaan type(HTN)and one strain of Seoul type(SEO).
Conclusion
In Tiantai County,Apodemus agrarius is the main source of HFRS infection;the main epidemic type of HV is HTN and SEO is first found in Apodemus agrarius.