By using the wastes fish skin of sturgeon processed as a raw material, a macromolecule biomaterial of collagen was extracted. Acid-soluble collagen(ASC) and pepsin-soluble collagen(PSC) were successfully isolated from...By using the wastes fish skin of sturgeon processed as a raw material, a macromolecule biomaterial of collagen was extracted. Acid-soluble collagen(ASC) and pepsin-soluble collagen(PSC) were successfully isolated from the skin of hybrid sturgeon with two extraction methods. The yields of ASC and PSC based on the wet weight of skin were 5.73 ± 0.11% and 10.26 ± 0.39%, respectively. The denaturation and melting points of ASC(26.83 ℃ and 110.49 ℃) and PSC(26.54 ℃ and 102.99 ℃) were assessed by Circular dichroism(CD) and Differential scanning calorimetry(DSC). ASC and PSC appeared to be dense sheet-like film linked by random-coiled filaments under scanning electron microscopy(SEM). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) and Fourier transform infrared spectroscopy(FTIR) confirmed that both the ASC and PSC were Type I collagen and maintained a complete triple helix structure. These results indicated that both ASC and PSC possessed good biological activity and could be widely used in medical biomaterials and other fields.展开更多
The aim of this research was to assess the diversity of the Cameroon cotton zone in soybean associated rhizobia in order to formulate the most efficient elite inoculant to boost both the cotton and soybean production....The aim of this research was to assess the diversity of the Cameroon cotton zone in soybean associated rhizobia in order to formulate the most efficient elite inoculant to boost both the cotton and soybean production. Therefore, soybean associated rhizobia were isolated and characterized morphologically, physiologically and biochemically on YEMA culture media. For each of the two soybean varieties (Houla1 and TGX1910 14F) used, the trials were laid out in two IRAD-fields of North Cameroon (Sanguere-Paul) and Far-North (Soukoundou) respectively, under a complete randomized complete block design, the isolate formulations representing the treatments. The six isolated strains (IS1, IS2, IS3, IS4, IS5, IS6) from which seven liquid inoculant were formulated were revealed to belong to the same slow growing group of rhizobia, with a high level of tolerance to temperature, pH, and salinity, with optimum growth at respectively 28˚C, pH (7 - 9), salt (1% - 5%). Not surprisingly, root nodules were formed by both inoculated and uninoculated soybean plants. However, the most efficient soybean-rhizobia symbiosis for nodulations were isolate IS6 associated to TGX1910 14F variety, and isolate IS5 associated to Houla1variety at Sanguere-Paul. Whereas isolate M was associated to TGX1910 14F variety, Houla 1 variety had affinity with native rhizobia isolates at Soukoundou. The present results suggest the adaptability of rhizobia isolates to a particular soybean variety at a particular cotton fields zone. These findings should be taken into consideration for commercial inoculant formulation.展开更多
Collagen of squid(Ommastrephes bartrami) skin was examined in the present study. Histology showed that collagen fiber in the skin was partially cross-linked with muscle fiber. Acid-solubilized collagen(ASC) and pepsin...Collagen of squid(Ommastrephes bartrami) skin was examined in the present study. Histology showed that collagen fiber in the skin was partially cross-linked with muscle fiber. Acid-solubilized collagen(ASC) and pepsin-solubilized collagen(PSC) were extracted from the skin and characterized. The results of amino acid composition and electrophoretic patterns revealed that ASC and PSC were both type Ⅰ collagen,containing α1 and α2 chains. FTIR(fourier transform infrared spectroscopy) investigations confirmed the existence of helical arrangements in PSC of squid skin. The denaturation temperature(Td) and shrinkage temperature(Ts) of PSC were 29.4℃ and 52.8℃,respectively.展开更多
A simple and rapid Supercritical Fluid Chromatography (SFC) method has been developed to isolate and characterize R-Isomer of Ezetimi be by using normal phase Chiral Cel OD-H with 250 mm × 30 mm, 5 microns column...A simple and rapid Supercritical Fluid Chromatography (SFC) method has been developed to isolate and characterize R-Isomer of Ezetimi be by using normal phase Chiral Cel OD-H with 250 mm × 30 mm, 5 microns column using a mobile phase system containing super critical fluid carbondi oxide (Co2) and the percentage of 2-Propanol as a mobile phase (85:15) and detection at 230 nm. The isolated R-Isomer is characterized by using UV-vis, FT-IR, ESI-MS, HPLC1H and 13C NMR. The purity of isolated R-Isomer is about 98%.展开更多
The analysis of virus genome is based on nucleic acid isolation. The aims of this study were to develop a method forisolation and identification of virus double-stranded ribonucleic acid (dsRNA) and to elucidate the n...The analysis of virus genome is based on nucleic acid isolation. The aims of this study were to develop a method forisolation and identification of virus double-stranded ribonucleic acid (dsRNA) and to elucidate the nucleotide sequencesof strawberry virus. Using the modified method, virus dsRNA was extracted from strawberry virus indicator plants andcultivated strawberry plants and detected using agarose gel electrophoresis with ethidium bromide staining and reversetranscription-polymerase chain reaction (RT-PCR). The quantity of virus dsRNA varied among strawberry cultivars. Thequantity of dsRNA from in vitro plantlets was higher than that from the young leaves of field plants. For the field-grownplants, there was more dsRNA in the young leaves. Virus dsRNA extracted from strawberry plants was resistant todeoxyribonuclease Ⅰ(DNase Ⅰ), but evidently, it became resistant to ribonuclease A (RNase A) only in the presence of0.5 M NaCl. Its bands in agarose gel could be readily recycled using an agarose gel DNA purification kit. With RT-PCR,the segments of both strawberry mottle virus and Strawberry mild yellow edge virus genomes were amplified by usingthe virus dsRNA recycled from gel or treated with DNase Ⅰ /RNase A as templates. The system developed for dsRNAisolation and identification in strawberry plants laid a sound foundation for the work on genome analysis of strawberryvirus isolates in China.展开更多
Tomato(Solanum lycopersicum) plants exhibiting severe leaf distortion, mottle and systemic crinkling symptoms were identified in Hainan province in China in 2016. To survey and control the disease, it is necessary to ...Tomato(Solanum lycopersicum) plants exhibiting severe leaf distortion, mottle and systemic crinkling symptoms were identified in Hainan province in China in 2016. To survey and control the disease, it is necessary to identify and characterize the pathogen causing the disease. Dot enzyme-linked immunosorbent assay showed that the crude saps of the infected tomato samples reacted positively with the monoclonal antibody against Tobacco mosaic virus which indicated that one or more tobamoviruses are likely associated with the disease. RT-p CR and DNA sequence analysis results further elucidated that Tomato mottle mosaic virus(To MMV) in Tobamovirus was the pathogen causing the mottle disease in tomato. We amplified and sequenced the full-length sequence of the genome which showed the highest nucleotide identity with To MMV YYMLJ and To MMV Ti Lha LJ isolates. The putative virus isolate was named To MMV Hainan. Biological indexing studies showed that To MMV Hainan can infect Nicotiana benthamiana, Capsicum annuum and Solanum lycopersicum showing serious symptoms. This was the first identification and characterization of To MMV infecting tomato in Hainan of China.展开更多
[Objective] The aim of this study was to determine and analyze the pathogen and the reasons of duck viral hepatitis which is prevalent recently and difficult to control.[Method] Viruses were isolated from livers and s...[Objective] The aim of this study was to determine and analyze the pathogen and the reasons of duck viral hepatitis which is prevalent recently and difficult to control.[Method] Viruses were isolated from livers and spleens of ducks with typical clinical symptoms in Linyi,Weifang,Binzhou and other regions of Shandong Province.The pathogenic characteristics were observed by inoculation in chicken or duck embryo,RT-PCR,serological test,and duck regression.[Result] Four duck hepatitis virus (DHV) strains were isolated,and the 5th passage allantoic fluid contained 103.41-105.20 ELD50/ml.The serum cross protection rate was 20%-80% between the DHV stains and DHV type I.The mortalities of 4-day-old healthy ducks challenged by these four stains were 50%-100%.All challenged ducks had typical lesions of duck viral hepatitis,and the death peak appeared after 24-48 h.[Conclusion] The virulence of different DHV isolates has regional difference.展开更多
In this study,the liver,kidney and spleen tissues were collected from pigs with suspected PR in a pig farm in Jiangyan District,Taizhou City for virus isolation and identification. The isolated virus was inoculated on...In this study,the liver,kidney and spleen tissues were collected from pigs with suspected PR in a pig farm in Jiangyan District,Taizhou City for virus isolation and identification. The isolated virus was inoculated onto PK15 monolayer cells. The virus culture was collected to extract genomic DNA for PCR assay and indirect immunoinfluscent assay. The results showed that the isolated virus was porcine pseudorabies virus,which was named TAIZ130417. The growth titer of the isolated virus reached 108. 12 TCID50/ml on PK15 cells. Rabbits inoculated with the isolated virus soon exhibited pseudorabies symptoms such as itching and eventually died. The results provided reference for in-depth research and scientific prevention and control of pseudorabies.展开更多
Duck tembusu virus disease is one of the most serious infectious diseases endangering duck industry. A strain of virus was isolated from a dead duck,and performed PCR identification and sequence analysis. The results ...Duck tembusu virus disease is one of the most serious infectious diseases endangering duck industry. A strain of virus was isolated from a dead duck,and performed PCR identification and sequence analysis. The results showed that the sequence of the isolate shared above 99% homology with duck tembusu virus( DTMUV) sequence on Gen Bank. The result indicated that the isolated virus was DTMUV.展开更多
Porcine epidemic diarrhea(PED)is caused by porcine epidemic diarrhea virus(PEDV),and is characterized by vomiting,diarrhea and dehydration of suckling pigs from 80% to 100% morbidity and 50% to 90% mortality,and resul...Porcine epidemic diarrhea(PED)is caused by porcine epidemic diarrhea virus(PEDV),and is characterized by vomiting,diarrhea and dehydration of suckling pigs from 80% to 100% morbidity and 50% to 90% mortality,and resulted in tremendous economic losses to swine industry.The PEDV mainly infects small intestine of pigs,resulting in vacuolar degeneration and necrosis of mucosal epithelium.The IPEC-J2 is a pig intestine epithelial cell line,which is similar to the intestinal environment of piglets,can be used to isolate and identify the PEDV field isolates.In this study,it appeared the PEDV typical postmortem changes and histopathological lesion of degeneration and destruction of small intestine in infected piglets,and IHC identified that the PEDV distributed in the mucosa and submucosa of small intestine mostly.Furthermore,the PEDV HLJ strain was successfully isolated and characterized in the IPEC-J2 cells,and indicated that the IPEC-J2 cell line was sensitive to isolate and adapt the PEDV field strain,and could be utilized to multiply the PEDV rapidly.The S gene analysis indicated that the PEDV HLJ strain was the prevailed virus,belonged to Group 1 with attenuated virulent DR13,SC1402 and J-S2/2015 strains isolated in South Korea and China from 2014 to 2015.This study had important theoretical and practical significances on analyzing genetic variation of the PEDV,understanding the pathogenic characteristics of the virus and developing new vaccines for the PED.展开更多
The present research work was carried out to isolate and identify Newcastle disease virus (NDV) by using haemagglutination inhibition (HI) test and HA-HI virus isolation, embryonated eggs (EE) and chicken embryo fibro...The present research work was carried out to isolate and identify Newcastle disease virus (NDV) by using haemagglutination inhibition (HI) test and HA-HI virus isolation, embryonated eggs (EE) and chicken embryo fibroblasts (CEF). A total of 95 clinical (blood, tracheal and cloacal swabs) and post-mortem (brain, lung, colon and spleen) samples were collected from chickens of field outbreaks of suspected Newcastle disease virus (NDV). The HI and HA-HI were employed to detect NDV in tissue homogenates of all the clinical and post-mortem samples as well as laboratory samples (AF and ICF). Among the four different types of post-mortem samples, virus isolation rate was found to be low in body organs. In CEF cell culture system, the rate of virus isolation from all the aforesaid samples was found to be at 100% with the exception of serum samples;while in tracheal and cloacal swabs, it was at 90%;while in serum, it was at 10%, in all clinical cases. The isolation rate of NDV was higher in CEF culture system (66.7%) compared to that of avian embryos (33.3%). Samples were inoculated and the allantoic fluid (AF) of the dead embryos and the infected culture fluid (ICF) of the CEF were harvested at 24 to 96 hours of the post-infection, respectively, which revealed that the virulent strain of NDV is highly prevalent in the region. The prevalence of NDV was established at 1.1%, 2.1% and 4.2% using HA-HI, EE, and CEF methods. Rapid detection and identification of the virus are crucial for the effective control of the disease as conventional diagnostic methods such as virus isolation on embryonated eggs followed by serological identification in haemagglutination-inhibition test are laborious and time-consuming. The speed of the diagnosis can be considerably increased by using methods based on molecular biology, e.g. reverse transcription—polymerase chain reaction. However, the genetic variability of APMV-1 isolates should be considered carefully as the potential cause for false negative results of genetic-based laboratory tests.展开更多
Encephalomyocarditis virus(EMCV)is a positive single-stranded small RNA virus without envelope,which can infect a variety of mammals.Swines are the most susceptible animals,which can cause acute myocarditis and respir...Encephalomyocarditis virus(EMCV)is a positive single-stranded small RNA virus without envelope,which can infect a variety of mammals.Swines are the most susceptible animals,which can cause acute myocarditis and respiratory failure in piglets and reproductive failure in pregnant sows.Diseases caused by EMCV have a wide range of effects on the global swine industry.In this study,a strain of EMCV was isolated from a swine aborted fetus in northeast China.It was identified by reverse transcriptase polymerase chain reaction(RT-PCR),electron microscopic observation and indirect immunofluorescence assay.The subsequent results showed that the virus titer of HLJ strain grew to 8.3 lgTCID50 on baby hamster kidney 21(BHK-21)cells.And HLJ strain caused the specific cytopathic effect(CPE)on BHK-21 cells and severe pathological changes in mice.Complete genome sequencing and multiple sequence alignment showed that the homology between HLJ strain and other isolates worldwide was 71.5%-99.7%.Phylogenetic analysis showed that EMCV isolates fell into five clusters:lineageⅠ,Ⅱ,Ⅲ,ⅣandⅤ,based on the nucleotide sequences of the entire open reading frame(ORF)and VP1 gene.HLJ isolate was grouped into lineage I.The analyses of amino acid mutation sites of VP1 protein showed that the amino acids at positions 20 and 54 in VP1 junction were unique to HLJ strain.The isolation of HLJ strain enriched the epidemiological database of EMCV.展开更多
Tristeza is a devastating viral disease in all the citrus growing countries throughout the world and has killed millions of citrus trees in severely affected orchards.The citrus species grafted on sour orange rootstoc...Tristeza is a devastating viral disease in all the citrus growing countries throughout the world and has killed millions of citrus trees in severely affected orchards.The citrus species grafted on sour orange rootstock are affected by this disease.Predominantly,the sweet orange,grapefruit and lime trees grafted on sour orange exhibit severe symptoms like quick decline,vein clearing,pin holing,bark scaling and degeneration leading to variable symptoms.Symptomatic expression of Citrus tristeza virus(CTV)in different hosts has been attributed to virus isolates which are from severe to mild.Different serological and molecular assays have been deployed to differentiate the strains of CTV.Citrus tristeza virus is diversified towards its strains on the basis of biological,serological and molecular characterization.Phenotypic expression is due to genetic alteration and different molecular basis have now been adopted for strain differentiation.This review will give a brief idea about the different CTV isolates,their characterization based on nucleic acid and serological assays.Different methods along with salient features for strain characterization has also been reviewed.This review will also open the new aspects towards formulation of management strategies through different detection techniques.展开更多
<span style="font-family:Verdana;">The present work encompasses identification and characterization of major degradation product (DP) of OSM observed in base hydrolytic stress study. The separation of ...<span style="font-family:Verdana;">The present work encompasses identification and characterization of major degradation product (DP) of OSM observed in base hydrolytic stress study. The separation of DP was carried out on a non-polar stationary phase by using high-performance liquid chromatography system (HPLC). Using waters X-bridge (250 mm × 4.6 mm, 5 μm) C18 column with gradient elution program. For the characterization study, stress samples were subjected to HPLC and UPLC-QTOF-MS/MS and based on mass fragmentation pattern</span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">,</span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;"> plausible structure was deduced. Further</span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">,</span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;"> the DP was isolated using semi-prepara</span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">- </span></span></span><span><span><span style="font-family:;" "=""><span style="font-family:Verdana;">tive liquid chromatography and concentrated the fractions using lyophiliza</span><span style="font-family:Verdana;">tion. The isolated DP was subjected to extensive 1D (1H, 13C, and</span><span style="font-family:Verdana;"> DEPT-135) and 2D (COSY, HSQC and HMBC) nuclear magnetic resonance (NMR) studies to authenticate the structure. The impurity was unambiguously named as N-(2-((2-(dimethylamino)ethyl)(methyl)amino)-4-metho</span></span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">-</span></span></span><span><span><span style="font-family:;" "=""><span style="font-family:Verdana;">xy-5-((4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl)amino)phenyl)-3-methoxy</span><span style="font-family:Verdana;">propanamide.</span></span></span></span><span><span><span style="font-family:;" "=""> </span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">Add</span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">- </span></span></span><span><span><span style="font-family:;" "=""><span style="font-family:Verdana;">itionally, the </span><i><span style="font-family:Verdana;">In-Silico</span></i><span style="font-family:Verdana;"> structure activity relation (QSAR) assessed through sta</span></span></span></span><span><span><span style="font-family:;" "=""><span style="font-family:Verdana;">tistical based software’s DEREK Nexus</span><sup><span style="font-family:Verdana;">TM</span></sup><span style="font-family:Verdana;">, and MultiCASE, Case Ultra</span><sup><span style="font-family:Verdana;">TM</span></sup></span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;"> widely accepted and respected software’s for DP and OSM</span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">.</span></span></span>展开更多
A novel antimicrobial polypeptide was isolated and characterized from loach, Misgurnus anguillicaudatus. The polypeptide, named MAPP, is a single-chain polypeptide with Mw about 9,800 Dalton and pI about 4.78; the N-t...A novel antimicrobial polypeptide was isolated and characterized from loach, Misgurnus anguillicaudatus. The polypeptide, named MAPP, is a single-chain polypeptide with Mw about 9,800 Dalton and pI about 4.78; the N-tag of MAPP was CFGWN. MAPP showed good inhibition against various bacteria including Bacillus subtilis, Escherichia coli and Staphylococcus aureus. MAPP could be used as a lead compound in antibiotics drug discovery.展开更多
A Novel stability indicating RP-UPLC chromatographic method was developed for analysis of Nevirapine in pharmaceutical formulations. The developed RP-UPLC method is superior in technology to conventional RP-HPLC with ...A Novel stability indicating RP-UPLC chromatographic method was developed for analysis of Nevirapine in pharmaceutical formulations. The developed RP-UPLC method is superior in technology to conventional RP-HPLC with respect to speed, resolution, solvent consumption and cost of analysis. Nevirapine was subjected to the stress conditions like acid, base, thermal, oxidative and photolytic degradation. Nevirapine was found to degrade significantly in acid and thermal degradation. In acid degradation relative retention time with 0.42 is found as unknown impurity. New impurity was identified, isolated using mass based auto purification system and characterized by <sup>1</sup>H NMR (<sup>1</sup>D and <sup>2</sup>D) and HRMS experiments. Isolated impurity was showing molecular weight of 244.10, molecular formula C<sub>12</sub>H<sub>12</sub>N<sub>4</sub>O<sub>2</sub> and its name as 2-(3-Amino-4-methylpyridin-2-ylamino)nicotinic acid. The calibration graph was linear and the method showed less deviation in accuracy results. The test solution was found to be stable for 20 days when stored in the refrigerator between 2°C to 8°C. The developed RP-UPLC method was validated and meets the requirements delineated by the International Conference on Harmonization (ICH) guidelines. The intra-day and inter-day variation was less than 1%. The method was reproducible and selective for the estimation of Nevirapine. Because the method could effectively separate the drug from its degradation products, it can be employed as a stability-indicating method.展开更多
Cytoplasmic male sterility(CMS) is a maternally inherited trait that results in the failure to produce functional pollen.It was identified in many plants,and it is widely used to exploit heterosis.
In November 2009, a respiratory disease with rapid transmission, rapid onset and mortality of about 8% appeared many times in a large chicken farm in Jiangsu Province of China. Necropsy revealed tracheal bleeding, kid...In November 2009, a respiratory disease with rapid transmission, rapid onset and mortality of about 8% appeared many times in a large chicken farm in Jiangsu Province of China. Necropsy revealed tracheal bleeding, kidney enlargement and white-spotted kidney. An isolate from the farm was identified as an avian infectious bronchitis virus (IBV) by chicken embryo inoculation, hemagglutination assay, virus interference assay, animal regression and tracheal rings culture. The complete S1 gene was amplified by RT-PCR, and its homology to that of the vaccine strains commonly used in China was analyzed with DNAStar software. Therefore, the IBV isolate was initially classified into nephropathogenic IBV and named IBV JS09 strain.展开更多
基金Funded by the National Natural Science Foundation of China(No.51173143)
文摘By using the wastes fish skin of sturgeon processed as a raw material, a macromolecule biomaterial of collagen was extracted. Acid-soluble collagen(ASC) and pepsin-soluble collagen(PSC) were successfully isolated from the skin of hybrid sturgeon with two extraction methods. The yields of ASC and PSC based on the wet weight of skin were 5.73 ± 0.11% and 10.26 ± 0.39%, respectively. The denaturation and melting points of ASC(26.83 ℃ and 110.49 ℃) and PSC(26.54 ℃ and 102.99 ℃) were assessed by Circular dichroism(CD) and Differential scanning calorimetry(DSC). ASC and PSC appeared to be dense sheet-like film linked by random-coiled filaments under scanning electron microscopy(SEM). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) and Fourier transform infrared spectroscopy(FTIR) confirmed that both the ASC and PSC were Type I collagen and maintained a complete triple helix structure. These results indicated that both ASC and PSC possessed good biological activity and could be widely used in medical biomaterials and other fields.
文摘The aim of this research was to assess the diversity of the Cameroon cotton zone in soybean associated rhizobia in order to formulate the most efficient elite inoculant to boost both the cotton and soybean production. Therefore, soybean associated rhizobia were isolated and characterized morphologically, physiologically and biochemically on YEMA culture media. For each of the two soybean varieties (Houla1 and TGX1910 14F) used, the trials were laid out in two IRAD-fields of North Cameroon (Sanguere-Paul) and Far-North (Soukoundou) respectively, under a complete randomized complete block design, the isolate formulations representing the treatments. The six isolated strains (IS1, IS2, IS3, IS4, IS5, IS6) from which seven liquid inoculant were formulated were revealed to belong to the same slow growing group of rhizobia, with a high level of tolerance to temperature, pH, and salinity, with optimum growth at respectively 28˚C, pH (7 - 9), salt (1% - 5%). Not surprisingly, root nodules were formed by both inoculated and uninoculated soybean plants. However, the most efficient soybean-rhizobia symbiosis for nodulations were isolate IS6 associated to TGX1910 14F variety, and isolate IS5 associated to Houla1variety at Sanguere-Paul. Whereas isolate M was associated to TGX1910 14F variety, Houla 1 variety had affinity with native rhizobia isolates at Soukoundou. The present results suggest the adaptability of rhizobia isolates to a particular soybean variety at a particular cotton fields zone. These findings should be taken into consideration for commercial inoculant formulation.
基金supported by the High Technology Research and Development Programme of China (No. 2006AA09Z438)the National Natural Science Foundation of China (No.30871943).
文摘Collagen of squid(Ommastrephes bartrami) skin was examined in the present study. Histology showed that collagen fiber in the skin was partially cross-linked with muscle fiber. Acid-solubilized collagen(ASC) and pepsin-solubilized collagen(PSC) were extracted from the skin and characterized. The results of amino acid composition and electrophoretic patterns revealed that ASC and PSC were both type Ⅰ collagen,containing α1 and α2 chains. FTIR(fourier transform infrared spectroscopy) investigations confirmed the existence of helical arrangements in PSC of squid skin. The denaturation temperature(Td) and shrinkage temperature(Ts) of PSC were 29.4℃ and 52.8℃,respectively.
文摘A simple and rapid Supercritical Fluid Chromatography (SFC) method has been developed to isolate and characterize R-Isomer of Ezetimi be by using normal phase Chiral Cel OD-H with 250 mm × 30 mm, 5 microns column using a mobile phase system containing super critical fluid carbondi oxide (Co2) and the percentage of 2-Propanol as a mobile phase (85:15) and detection at 230 nm. The isolated R-Isomer is characterized by using UV-vis, FT-IR, ESI-MS, HPLC1H and 13C NMR. The purity of isolated R-Isomer is about 98%.
基金supported by a grant from the National Natural Science Foundation of China(30200187)a grant from the Scientific Research Start-up Foundation of Ministry of Education for Returned 0verseas Students,China(2002-247).
文摘The analysis of virus genome is based on nucleic acid isolation. The aims of this study were to develop a method forisolation and identification of virus double-stranded ribonucleic acid (dsRNA) and to elucidate the nucleotide sequencesof strawberry virus. Using the modified method, virus dsRNA was extracted from strawberry virus indicator plants andcultivated strawberry plants and detected using agarose gel electrophoresis with ethidium bromide staining and reversetranscription-polymerase chain reaction (RT-PCR). The quantity of virus dsRNA varied among strawberry cultivars. Thequantity of dsRNA from in vitro plantlets was higher than that from the young leaves of field plants. For the field-grownplants, there was more dsRNA in the young leaves. Virus dsRNA extracted from strawberry plants was resistant todeoxyribonuclease Ⅰ(DNase Ⅰ), but evidently, it became resistant to ribonuclease A (RNase A) only in the presence of0.5 M NaCl. Its bands in agarose gel could be readily recycled using an agarose gel DNA purification kit. With RT-PCR,the segments of both strawberry mottle virus and Strawberry mild yellow edge virus genomes were amplified by usingthe virus dsRNA recycled from gel or treated with DNase Ⅰ /RNase A as templates. The system developed for dsRNAisolation and identification in strawberry plants laid a sound foundation for the work on genome analysis of strawberryvirus isolates in China.
基金supported by the Agricultural Science and Technology Innovation program, China (ASTIp)
文摘Tomato(Solanum lycopersicum) plants exhibiting severe leaf distortion, mottle and systemic crinkling symptoms were identified in Hainan province in China in 2016. To survey and control the disease, it is necessary to identify and characterize the pathogen causing the disease. Dot enzyme-linked immunosorbent assay showed that the crude saps of the infected tomato samples reacted positively with the monoclonal antibody against Tobacco mosaic virus which indicated that one or more tobamoviruses are likely associated with the disease. RT-p CR and DNA sequence analysis results further elucidated that Tomato mottle mosaic virus(To MMV) in Tobamovirus was the pathogen causing the mottle disease in tomato. We amplified and sequenced the full-length sequence of the genome which showed the highest nucleotide identity with To MMV YYMLJ and To MMV Ti Lha LJ isolates. The putative virus isolate was named To MMV Hainan. Biological indexing studies showed that To MMV Hainan can infect Nicotiana benthamiana, Capsicum annuum and Solanum lycopersicum showing serious symptoms. This was the first identification and characterization of To MMV infecting tomato in Hainan of China.
基金funded by the Technology Development Program of Chengyang District of Qingdao City (2008-4-sf)
文摘[Objective] The aim of this study was to determine and analyze the pathogen and the reasons of duck viral hepatitis which is prevalent recently and difficult to control.[Method] Viruses were isolated from livers and spleens of ducks with typical clinical symptoms in Linyi,Weifang,Binzhou and other regions of Shandong Province.The pathogenic characteristics were observed by inoculation in chicken or duck embryo,RT-PCR,serological test,and duck regression.[Result] Four duck hepatitis virus (DHV) strains were isolated,and the 5th passage allantoic fluid contained 103.41-105.20 ELD50/ml.The serum cross protection rate was 20%-80% between the DHV stains and DHV type I.The mortalities of 4-day-old healthy ducks challenged by these four stains were 50%-100%.All challenged ducks had typical lesions of duck viral hepatitis,and the death peak appeared after 24-48 h.[Conclusion] The virulence of different DHV isolates has regional difference.
基金Supported by Youth Fund of Jiangsu Agri-animal Husbandry Vocational College(NSFQN1304)Key Project of Jiangsu Agri-animal Husbandry Vocational College(ZD201104)Phoenix Talent Project of Jiangsu Agri-animal Husbandry Vocational College(10434014001)
文摘In this study,the liver,kidney and spleen tissues were collected from pigs with suspected PR in a pig farm in Jiangyan District,Taizhou City for virus isolation and identification. The isolated virus was inoculated onto PK15 monolayer cells. The virus culture was collected to extract genomic DNA for PCR assay and indirect immunoinfluscent assay. The results showed that the isolated virus was porcine pseudorabies virus,which was named TAIZ130417. The growth titer of the isolated virus reached 108. 12 TCID50/ml on PK15 cells. Rabbits inoculated with the isolated virus soon exhibited pseudorabies symptoms such as itching and eventually died. The results provided reference for in-depth research and scientific prevention and control of pseudorabies.
基金Supported by National Natural Science Foundation of China(31402224)Key Research & Development Project of Hainan Province(ZDYF2017029)Agricultural Science and Technology Innovation Project of Hainan Academy of Agricultural Sciences(CXZX201413)
文摘Duck tembusu virus disease is one of the most serious infectious diseases endangering duck industry. A strain of virus was isolated from a dead duck,and performed PCR identification and sequence analysis. The results showed that the sequence of the isolate shared above 99% homology with duck tembusu virus( DTMUV) sequence on Gen Bank. The result indicated that the isolated virus was DTMUV.
基金Supported by the National Natural Science Foundation of China(31201911 31372438)
文摘Porcine epidemic diarrhea(PED)is caused by porcine epidemic diarrhea virus(PEDV),and is characterized by vomiting,diarrhea and dehydration of suckling pigs from 80% to 100% morbidity and 50% to 90% mortality,and resulted in tremendous economic losses to swine industry.The PEDV mainly infects small intestine of pigs,resulting in vacuolar degeneration and necrosis of mucosal epithelium.The IPEC-J2 is a pig intestine epithelial cell line,which is similar to the intestinal environment of piglets,can be used to isolate and identify the PEDV field isolates.In this study,it appeared the PEDV typical postmortem changes and histopathological lesion of degeneration and destruction of small intestine in infected piglets,and IHC identified that the PEDV distributed in the mucosa and submucosa of small intestine mostly.Furthermore,the PEDV HLJ strain was successfully isolated and characterized in the IPEC-J2 cells,and indicated that the IPEC-J2 cell line was sensitive to isolate and adapt the PEDV field strain,and could be utilized to multiply the PEDV rapidly.The S gene analysis indicated that the PEDV HLJ strain was the prevailed virus,belonged to Group 1 with attenuated virulent DR13,SC1402 and J-S2/2015 strains isolated in South Korea and China from 2014 to 2015.This study had important theoretical and practical significances on analyzing genetic variation of the PEDV,understanding the pathogenic characteristics of the virus and developing new vaccines for the PED.
文摘The present research work was carried out to isolate and identify Newcastle disease virus (NDV) by using haemagglutination inhibition (HI) test and HA-HI virus isolation, embryonated eggs (EE) and chicken embryo fibroblasts (CEF). A total of 95 clinical (blood, tracheal and cloacal swabs) and post-mortem (brain, lung, colon and spleen) samples were collected from chickens of field outbreaks of suspected Newcastle disease virus (NDV). The HI and HA-HI were employed to detect NDV in tissue homogenates of all the clinical and post-mortem samples as well as laboratory samples (AF and ICF). Among the four different types of post-mortem samples, virus isolation rate was found to be low in body organs. In CEF cell culture system, the rate of virus isolation from all the aforesaid samples was found to be at 100% with the exception of serum samples;while in tracheal and cloacal swabs, it was at 90%;while in serum, it was at 10%, in all clinical cases. The isolation rate of NDV was higher in CEF culture system (66.7%) compared to that of avian embryos (33.3%). Samples were inoculated and the allantoic fluid (AF) of the dead embryos and the infected culture fluid (ICF) of the CEF were harvested at 24 to 96 hours of the post-infection, respectively, which revealed that the virulent strain of NDV is highly prevalent in the region. The prevalence of NDV was established at 1.1%, 2.1% and 4.2% using HA-HI, EE, and CEF methods. Rapid detection and identification of the virus are crucial for the effective control of the disease as conventional diagnostic methods such as virus isolation on embryonated eggs followed by serological identification in haemagglutination-inhibition test are laborious and time-consuming. The speed of the diagnosis can be considerably increased by using methods based on molecular biology, e.g. reverse transcription—polymerase chain reaction. However, the genetic variability of APMV-1 isolates should be considered carefully as the potential cause for false negative results of genetic-based laboratory tests.
基金Supported by the National Natural Science Foundation of China(31372438)。
文摘Encephalomyocarditis virus(EMCV)is a positive single-stranded small RNA virus without envelope,which can infect a variety of mammals.Swines are the most susceptible animals,which can cause acute myocarditis and respiratory failure in piglets and reproductive failure in pregnant sows.Diseases caused by EMCV have a wide range of effects on the global swine industry.In this study,a strain of EMCV was isolated from a swine aborted fetus in northeast China.It was identified by reverse transcriptase polymerase chain reaction(RT-PCR),electron microscopic observation and indirect immunofluorescence assay.The subsequent results showed that the virus titer of HLJ strain grew to 8.3 lgTCID50 on baby hamster kidney 21(BHK-21)cells.And HLJ strain caused the specific cytopathic effect(CPE)on BHK-21 cells and severe pathological changes in mice.Complete genome sequencing and multiple sequence alignment showed that the homology between HLJ strain and other isolates worldwide was 71.5%-99.7%.Phylogenetic analysis showed that EMCV isolates fell into five clusters:lineageⅠ,Ⅱ,Ⅲ,ⅣandⅤ,based on the nucleotide sequences of the entire open reading frame(ORF)and VP1 gene.HLJ isolate was grouped into lineage I.The analyses of amino acid mutation sites of VP1 protein showed that the amino acids at positions 20 and 54 in VP1 junction were unique to HLJ strain.The isolation of HLJ strain enriched the epidemiological database of EMCV.
文摘Tristeza is a devastating viral disease in all the citrus growing countries throughout the world and has killed millions of citrus trees in severely affected orchards.The citrus species grafted on sour orange rootstock are affected by this disease.Predominantly,the sweet orange,grapefruit and lime trees grafted on sour orange exhibit severe symptoms like quick decline,vein clearing,pin holing,bark scaling and degeneration leading to variable symptoms.Symptomatic expression of Citrus tristeza virus(CTV)in different hosts has been attributed to virus isolates which are from severe to mild.Different serological and molecular assays have been deployed to differentiate the strains of CTV.Citrus tristeza virus is diversified towards its strains on the basis of biological,serological and molecular characterization.Phenotypic expression is due to genetic alteration and different molecular basis have now been adopted for strain differentiation.This review will give a brief idea about the different CTV isolates,their characterization based on nucleic acid and serological assays.Different methods along with salient features for strain characterization has also been reviewed.This review will also open the new aspects towards formulation of management strategies through different detection techniques.
文摘<span style="font-family:Verdana;">The present work encompasses identification and characterization of major degradation product (DP) of OSM observed in base hydrolytic stress study. The separation of DP was carried out on a non-polar stationary phase by using high-performance liquid chromatography system (HPLC). Using waters X-bridge (250 mm × 4.6 mm, 5 μm) C18 column with gradient elution program. For the characterization study, stress samples were subjected to HPLC and UPLC-QTOF-MS/MS and based on mass fragmentation pattern</span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">,</span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;"> plausible structure was deduced. Further</span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">,</span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;"> the DP was isolated using semi-prepara</span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">- </span></span></span><span><span><span style="font-family:;" "=""><span style="font-family:Verdana;">tive liquid chromatography and concentrated the fractions using lyophiliza</span><span style="font-family:Verdana;">tion. The isolated DP was subjected to extensive 1D (1H, 13C, and</span><span style="font-family:Verdana;"> DEPT-135) and 2D (COSY, HSQC and HMBC) nuclear magnetic resonance (NMR) studies to authenticate the structure. The impurity was unambiguously named as N-(2-((2-(dimethylamino)ethyl)(methyl)amino)-4-metho</span></span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">-</span></span></span><span><span><span style="font-family:;" "=""><span style="font-family:Verdana;">xy-5-((4-(1-methyl-1H-indol-3-yl)pyrimidin-2-yl)amino)phenyl)-3-methoxy</span><span style="font-family:Verdana;">propanamide.</span></span></span></span><span><span><span style="font-family:;" "=""> </span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">Add</span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">- </span></span></span><span><span><span style="font-family:;" "=""><span style="font-family:Verdana;">itionally, the </span><i><span style="font-family:Verdana;">In-Silico</span></i><span style="font-family:Verdana;"> structure activity relation (QSAR) assessed through sta</span></span></span></span><span><span><span style="font-family:;" "=""><span style="font-family:Verdana;">tistical based software’s DEREK Nexus</span><sup><span style="font-family:Verdana;">TM</span></sup><span style="font-family:Verdana;">, and MultiCASE, Case Ultra</span><sup><span style="font-family:Verdana;">TM</span></sup></span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;"> widely accepted and respected software’s for DP and OSM</span></span></span><span style="font-family:Verdana;"><span style="font-family:Verdana;"><span style="font-family:Verdana;">.</span></span></span>
文摘A novel antimicrobial polypeptide was isolated and characterized from loach, Misgurnus anguillicaudatus. The polypeptide, named MAPP, is a single-chain polypeptide with Mw about 9,800 Dalton and pI about 4.78; the N-tag of MAPP was CFGWN. MAPP showed good inhibition against various bacteria including Bacillus subtilis, Escherichia coli and Staphylococcus aureus. MAPP could be used as a lead compound in antibiotics drug discovery.
文摘A Novel stability indicating RP-UPLC chromatographic method was developed for analysis of Nevirapine in pharmaceutical formulations. The developed RP-UPLC method is superior in technology to conventional RP-HPLC with respect to speed, resolution, solvent consumption and cost of analysis. Nevirapine was subjected to the stress conditions like acid, base, thermal, oxidative and photolytic degradation. Nevirapine was found to degrade significantly in acid and thermal degradation. In acid degradation relative retention time with 0.42 is found as unknown impurity. New impurity was identified, isolated using mass based auto purification system and characterized by <sup>1</sup>H NMR (<sup>1</sup>D and <sup>2</sup>D) and HRMS experiments. Isolated impurity was showing molecular weight of 244.10, molecular formula C<sub>12</sub>H<sub>12</sub>N<sub>4</sub>O<sub>2</sub> and its name as 2-(3-Amino-4-methylpyridin-2-ylamino)nicotinic acid. The calibration graph was linear and the method showed less deviation in accuracy results. The test solution was found to be stable for 20 days when stored in the refrigerator between 2°C to 8°C. The developed RP-UPLC method was validated and meets the requirements delineated by the International Conference on Harmonization (ICH) guidelines. The intra-day and inter-day variation was less than 1%. The method was reproducible and selective for the estimation of Nevirapine. Because the method could effectively separate the drug from its degradation products, it can be employed as a stability-indicating method.
文摘Cytoplasmic male sterility(CMS) is a maternally inherited trait that results in the failure to produce functional pollen.It was identified in many plants,and it is widely used to exploit heterosis.
文摘In November 2009, a respiratory disease with rapid transmission, rapid onset and mortality of about 8% appeared many times in a large chicken farm in Jiangsu Province of China. Necropsy revealed tracheal bleeding, kidney enlargement and white-spotted kidney. An isolate from the farm was identified as an avian infectious bronchitis virus (IBV) by chicken embryo inoculation, hemagglutination assay, virus interference assay, animal regression and tracheal rings culture. The complete S1 gene was amplified by RT-PCR, and its homology to that of the vaccine strains commonly used in China was analyzed with DNAStar software. Therefore, the IBV isolate was initially classified into nephropathogenic IBV and named IBV JS09 strain.
