1.Molecular Cloning and Nucleotide Sequence Analysis of th e M Genome Segment of Hantavirus Z37, a Chinese HFRS Vaccine Strain
Han-ping, ZHU ; He-bin, LIU ; Ping-ping, YAO ; Zhi-yong, ZHU
Virologica Sinica 2001;16(1):15-21
One of the strain of bivalent HFRS vaccine, Z37 strain was isolated from Rattus norvegicus and identified as SEO virus by serological test. The M segment cDNA of Hantavirus Z37 strain was obtained by reverse transcripti on and polymerase chain reaction, subsequently cloned into pGEM-T vector. The s equence of positive recombinants was determined by the method of dideoxy chain t ermination, which revealed that the M genomic segment is 3651 nucleotide in len gth with a predicated long open reading frame encoding a protein of 1133 amino acids. Comparison with HNT type (76-118, A9, HV-114 strains) indicated that th ere were 71.8%~72.1% homology at the nucleotide level, 76.2~76.7% homology at the amino acid level. Comparison with SEO type (R22,L99,80-39 strains) showed 95.3~96.1 homology at the nucleotide level, 95.3~99.2% homology at the amino acid level. The results of nucleotide and amino acid comparison indicated that Z 37 strain is SEO viruses in molecular level.
2.Advances in actinobacterial proteomics.
Yao ZHANG ; Ping XU ; Wenjun LI ; Yong TAO
Chinese Journal of Biotechnology 2014;30(7):1044-1058
Protein is the executor of physiological function, and direct embodiment of the life phenomena. Proteomics aims to systematically clarify all or parts of proteins' role and function in life movement. In post genome era, proteomics began to play more important role in life science field. Actinobacteria are closely linked to human production and life, which have produced many clinically important secondary metabolites, including antibiotics, antitumorals and enzymes. Actinobacterial systematics and its model organism Streptomyces coelicolor in 2001 genome sequence laid the foundation for further functional genomic studies. Actinobacterial proteomics was more directly and exactly to interpret the activity of life than genomics and transcriptomics, which grew much faster and received so much attention from scientists in the near years. Complex morphological differention, stronge environment adaptiveness, nitrogen-fixing capacity, metabolic mechanism, pathogenicity and natural produces' discovery were systematically reviewed in this study, which was expected to be the basis for promoting Actinobacterial proteomics study in the near future.
Actinobacteria
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genetics
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metabolism
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Genomics
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Proteomics
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Streptomyces coelicolor
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genetics
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metabolism
3.Expressions of NOS and NOS mRNA in the Lung of Rats with Hepatopulmonary Syndrome
xing-zhi, NI ; zhi-yong, WU ; zhi-ping, CHEN ; yao-lin, KUANG
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(08):-
Objective To investigate the expressions of nitric oxide synthase (NOS) protein and mRNA in the lung of rats with hepatopulmonary syndrome. Methods Male Sprague-Dawley rats were divided into four groups: sham operation (SO), intrahepatic portal hypertension (IHPH), prehepatic portal hypertension (PHPH) and portasymstimic shunt (PCS). Two weeks after preparation of rat models, the following measurements were performed: arterial blood gas analysis; the concentrations of NO in lungs; in situ hybridization of ecNOS and iNOS mRNA expressions in lung tissue sections with digoxin-labeled ecNOS and iNOS oligonucleotide probes; expressions of ecNOS and iNOS proteins by immunohistochemisty; image and semiquantitative analysis of the expressions of ecNOS, iNOS and their mRNA. Results PaO_ 2 was (73.85?6.51) mmHg in IHPH rats, significantly lower than that in PHPH, PCS and SO rats97.39? 1.33, 95.23?2.22 and (99.05?0.75)mmHg, respectively.The level of lung NO of IHPH was(19.78?5.33)?mol per gram of protein,much higher than that of PHPH, PCS and SO 13.21?3.99,13.89?3.16 and (8.71?1.68)?mol per gram of protein,respectively. In capillary endothelia, positive expressions of ecNOS mRNA and ecNOS protein in IHPH(4.96?0.82,4.11?0.28) were significantly higher than those of PHPH (1.81? 0.39, 1.63?0.18), PCS (1.88?0.53,1.83?0.16)and SO(1.19?0.32,0.98?0.20). Conclusion The expressions of NOS protein and mRNA in the lung of rats with hepatopulmonary syndrome were increased, and the level of lung NO was elevated, which seems to play an important role in the pathogenesis of hepatopulmonary syndrome.
4.Effect of matrine on NO and ADMA metabolism pathways in serum and tissues of mice with lipopolysaccharide-induced intestine tissue inflammation.
Yang WU ; Yang WANG ; Yao ZHANG ; Li-Ping CHEN ; Ji-Yong WANG
China Journal of Chinese Materia Medica 2014;39(12):2318-2321
OBJECTIVETo discuss the effect of matrine on nitric oxide (NO) and asymmetric methylarginine (ADMA) metabolism pathways in serum and tissues of mice with lipopolysaccharide (LPS) -induced intestine tissue inflammation.
METHODKunming mice were randomly divided into five groups: the normal control group, the LPS group and matrine (80, 40, 20 mg x kg(-1) x d(-1)) groups. The mice were intragastrically administered with drugs for 3 d (distilled water of the same volume for the normal control group and the LPS group). One hour after the last intragastrical administration, normal saline or LPS (1 mg x kg(-1)) were intraperitoneally injected. Twelve hours later, serum and tissues were collected to determine NO and ADMA levels and observe the pathological changes of intestinal tissues. The Western blot method was adopted to detect the protein expressions of arginine methyltransferases 1 (PRMT1) and dimethylarginine dimethylaminohydrolase 2 (DDAH2) in intestinal tissues.
RESULTCompared with the model group, matrine (80, 40, 20 mg x kg(-1) x d(-1)) groups showed lower NO content in serum and tissues, higher ADMA level in serum and increased PRMT1 expression in intestinal tissues, but without effect on DDAH2 expression.
CONCLUSIONMatrine could inhibit LPS-induced intestine tissue inflammation in mice. Its action mechanism is related to the decreased NO content in serum and tissues and increased ADMA level in serum and PRMT1 expression in intestinal tissues.
Alkaloids ; administration & dosage ; Animals ; Arginine ; analogs & derivatives ; blood ; metabolism ; Humans ; Inflammation ; Intestinal Diseases ; drug therapy ; enzymology ; immunology ; metabolism ; Intestines ; drug effects ; enzymology ; immunology ; metabolism ; Lipopolysaccharides ; adverse effects ; Male ; Mice ; Nitric Oxide ; blood ; metabolism ; Protein-Arginine N-Methyltransferases ; genetics ; metabolism ; Quinolizines ; administration & dosage
5.Effect of different water-improving methods on dental fluorosis of children in 50 drinking-water-borne fluorosis areas in Xi'an City
Lu DONG ; Wei CHEN ; Peijie YAO ; Xuehua SHI ; Shanshan HE ; Ping LI ; Yong WANG
Chinese Journal of Endemiology 2021;40(1):36-39
Objective:To evaluate the effect of different water-improving methods on dental fluorosis of children aged 8 to 12 in drinking-water-borne fluorosis areas in Xi'an City, and provide basis for making fine prevention and control measures.Methods:In 2018, 50 drinking-water-borne fluorosis villages in Xi'an City with water improvement time from 2011 to 2013 were selected as survey sites. The condition and the way of water improvement were surveyed, water fluoride content was determined and the prevalence of dental fluorosis in children aged 8 to 12 was examined, and the rates of dental fluorosis were compared before and after the water improvement in the 50 villages.Results:The water-improving projects were in normal operation in 50 villages. The medians of water fluoride content were 0.31, 0.88 and 1.14 mg/L in villages with municipal water supply, low fluorine well and defluoridation treatment water supply (the villages of different water-improvement methods were 12, 24 and 14, respectively), the differences were statistically significant ( H = 75.54, P < 0.01). The qualification rates of water fluoride in villages with different water-improvement methods were 100.00% (12/12), 79.17% (19/24) and 57.14% (8/14), and the difference was statistically significant (χ 2 = 6.95, P < 0.05). The detection rate of dental fluorosis was 43.17% (218/505) in the 11 villages with excessive water fluoride content, and the detection rate was 20.77% (331/1 594) in the 39 villages with qualified water fluoride content, the difference was statistically significant (χ 2 = 99.66, P < 0.01). After water improvement, the total detection rate of dental fluorosis (26.16%, 549/2 099) in the 50 villages was lower than that before water improvement (41.66%, 959/2 302), the difference was statistically significant (χ 2 = 117.17, P < 0.01). The detection rates of dental fluorosis were reduced after the water improvement in villages with municipal water supply and low fluorine well, the differences were statistically significant (χ 2 = 74.37, 69.36, P < 0.01). The detection rate was declined after water improvement in villages with defluoridation treatment water supply, but the difference was not significant (χ 2 = 0.78, P > 0.05). There was a statistically significant difference in the detection rate of dental fluorosis among children in villages with different water-improvement methods (χ 2 = 72.79, P < 0.01). The detection rate of dental fluorosis in villages with defluoridation treatment water supply [39.53% (200/506)] was higher than that in villages with municipal water supply [17.97% (133/740)] and low fluorine well [25.32% (216/853)], the differences were statistically significant ( P < 0.017); the detection rate of dental fluorosis in villages with municipal water supply was lower than that in villages with low fluorine well ( P < 0.017). Conclusions:The dental fluorosis of children's has been effectively controlled in the villages after water improvement in Xi'an City. The fluoride content in the water and the detection rate of dental fluorosis in children in some endemic areas are still high. It is necessary to further improve the water quality or consolidate the improvement effect as soon as possible.
6.Changes of Intestinal Flora of Children with Pneumonia Detected by Fluorescent Quantitative 16S rRNA/DNA Targeted Polymerase Chain Reaction Technique
yan-li, WANG ; hua, LIU ; ping, YAO ; ping, LU ; jing, XU ; li-fang, ZHOU ; yong-kun, HUANG
Journal of Applied Clinical Pediatrics 2003;0(10):-
Objective To study the intestinal flora changes of children with pneumonia,and explore the feasibility and practicability of fluorescent quantitative 16S rRNA/DNA targeted polymerase chain reaction(PCR) in quantitative study of bacterium.Methods The bacterial DNA was extracted in stool between 23 healthy children (control group)and 23 children (pneumonia group)with pneumonia after therapy,A260 of bacteria was detected and compared between 2 groups.16S rRNA/DNA PCR were applied to analyze and compare the bacterial content of lactobacilli and Escherichia coli in stool between control group and pneumonia group.Results Bacterial A260 of stool were respectively(3 381.2?817.2)mg/L in control group,(1 643.5?498.4)mg/L on therapy group post-treatment first day,(859.6?165.2) mg/ L on the third day of post-treatment,(1 263.8?337.3)mg/ L on the 7th day.There were significant differences of bacterial A260 between control group and pneumonia group in post-therapy(Pa
8.A clinical and pathologic study of meibomian gland carcinoma with reduplicative operations
Xiang-hua, WU ; Yong-ping, LI ; Ping, ZHANG ; Wen-xin, ZHANG ; Jian-xian, LIN ; Jian-liang, ZHENG ; Juan, YAO
Chinese Journal of Experimental Ophthalmology 2012;30(5):437-440
BackgroundThe meibomian gland carcinoma is an eyelid malignant tumor with a domestic incidence after basal cell carcinoma.Meibomian gland carcinoma is not sensitive to radiation therapy and chemotherapy,and the related factors with its recurrence and metastasis are rarely reported.ObjectiveThis study was to investigate the clinical and pathologic features of meibomian gland carcinoma with multiple operations and the effectiveness of histologically controlled excision.MethodsThe clinical data and the histopathologic sections of 34 cases of the meibomian gland carcinoma diagnosed by pathology were retrospectively analyzed at Zhongshan Ophthalmic Center in September 2003 to April 2011,and the treating effectiveness of histologically controlled excision was evaluated. ResultsIn this group of cases,the appearing rate of the meibomian gland carcinoma was resemble in both lateral eyes.A higher morbidity was on the upper eyelid (26/34,76.5%) and then the lower eyelids (5/34,14.7% ) and both (3/34,8.8%).The average ages of these cases were 57.5 years old.Sixteen of 21 misdiagnosed cases were identified as chalazion at the first visit,and no histopathological examination was performed in 11 cases after initial operation.Twenty-six cases(76.5% )were identified as meibomian gland carcinoma in initial histopathologic diagnosis.Two cases had histologically controlled excision and 16 cases had simple excision while 16 cases had chalazion enucleation in the first operation.All the patients had histologically controlled excision in Zhongshan Ophthalmic Center with 58.8% of the patients having pagetoid invasion.Sixteen cases were followed up for 5 months to 8 years after histologically controlled excision,in which none died of recurrence and metastasis of meibomian gland carcinoma.No significant differences were found in the pathological feature between 16 lost patients and 18 followed-up patients(P > 0.05 ).Conclusions Misdiagnosis of meibomian gland as chalazion is a main cause of repeat operations of meibomian gland carcinoma.Histologically controlled excision is a feasible therapy for the recurrence and metastasis of meibomian gland carcinoma.
9.Complete genome sequence analysis of Japanese encephalitis virus newly isolated in China.
Rong-Hui XIE ; Han-Ping ZHU ; Shi-Hong FU ; Yin-Kai CHENG ; Fang XU ; Ping-Ping YAO ; Zhang-Nv YANG ; Xiao-Long ZHOU ; Zhi-Yong ZHU
Chinese Journal of Experimental and Clinical Virology 2009;23(4):245-247
OBJECTIVETo study the complete genome sequence of Japanese encephalitis virus (JEV) strain XJ69 isolated in ZheJiang province and explore its evolution.
METHODSOverlapping primers were designed according to the full-length genomes from GenBank. RT-PCR was used to amplify the fragments and RT-PCR products were cloned T vector, sequenced and analyzed.
RESULTSThe genome of strain XJ69 and XJP613 were 10 964 nucleotides in length with a single open reading frame encoding 3432 amino acids. Comparison of the complete genome sequences of different JEV isolates showed XJ69 and XJP613 were 83.5%-99.2% and 83.4%-99.4% nucleotide sequence homology among them respectively, which resulted in 94.8%-99.7% amino acid sequence homology. Phylogenetic analysis through PrM/C,E and full-length genome showed that the XJ69 and XJP613 strain belonged to genotype I.
CONCLUSIONThe nucleotitede sequence and deduced amino acid sequence of XJ69 and XJP613 strain were similar to that of those of genotype I of Japanese encephalitis virus. It belonged to genotype I and were close to the isolates SH17M-07.
Animals ; Cell Line ; China ; Cricetinae ; Encephalitis Virus, Japanese ; classification ; genetics ; isolation & purification ; Encephalitis, Japanese ; virology ; Genome, Viral ; Humans ; Molecular Sequence Data ; Phylogeny
10.Molecular evolution analysis of hantaviruses in Zhejiang Province.
Ping-Ping YAO ; Han-Ping ZHU ; Xiao-Zhao DENG ; Fang XU ; Rong-Hui XIE ; Chen-Hui YAO ; Jing-Qing WENG ; Yun ZHANG ; Zhan-Qiu YANG ; Zhi-Yong ZHU
Chinese Journal of Virology 2010;26(6):465-470
In order to analyze the molecular epidemiology of Hantavirus (HV) in Zhejiang Province, the complete M and S genome sequences of 12 HV strains from different hosts and locations in Zhejiang Province of China during the period of 1981-2007 were analyzed on genetic evolution by DNAstar and MEGA 4.0 software in this research. Phylogenetic analyses revealed that HTN and SEO strains were co-circulating in Zhejiang Province, and the difference in sequence similarity and the phylogeny was closely related to the isolated regions, but had no distinct relationship with the isolate year and the host, indicating a relationship between epidemiology of HFRS and the distribution region, especially in HTNV. The isolates in the same region could be assigned in same or near phylogenetic clade sharing high sequence similarity. Interestingly, the Gou3 strain and ZJ5 strain isolated from Jiande region in Zhejiang Province formed a distinct phylogenetic lineage in SEOV clade, and different from the other SEOV variants outside China. We believed that the special SEOV variants were distributed in Jiande region.
Animals
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China
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Disease Reservoirs
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virology
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Evolution, Molecular
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Hantavirus
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classification
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genetics
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isolation & purification
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Hantavirus Infections
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virology
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Humans
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Molecular Sequence Data
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Phylogeny
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Rodentia
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virology
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Viral Proteins
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genetics