In order to investigate the prevalence and track genetic and antigenic evolutions of infectious bronchitis virus (IBV) and their prevalence in Guangxi, China since 1985, gene amplification and sequencing and virus neutralization (VN) test on chicken embryo tracheal organ cultures were used in genotyping and serotyping of 28 IBV isolates during 2009-2011 in Guangxi. The results of N gene sequencing and comparison showed that the 28 isolates and reference strains were classified into three groups, and most isolates belonged to group Ill, while the isolates in 1985-2008 belonged to groups IV and II. The data of VN test indicated that the 28 isolates belonged to 6 serotypes; among them, 71. 4% belonged to serotypes 1, 2, and 3, and 11 (39.3%) shared the same serotype with the current vaccine strains. Given the data of our previous study, it is found that prevalent serotypes and their proportions varied in different areas of Guangxi and during different periods. These data lay a good foundation for developing an oil-emulsified inactivated polyvalent vaccine containing local dominant serotypes for the effective prevention and control of infectious bronchitis.
Animals ; Antibodies, Viral ; immunology ; Chick Embryo ; Chickens ; China ; epidemiology ; Coronavirus Infections ; epidemiology ; immunology ; veterinary ; virology ; Infectious bronchitis virus ; classification ; genetics ; immunology ; isolation & purification ; Molecular Sequence Data ; Phylogeny ; Poultry Diseases ; epidemiology ; immunology ; virology

Monovalent antisera of 3 vaccine strains and 7 representative field isolates were prepared based on the comparison of genetic diversity of the hypervariable region I of S1 gene (HVR I from 3 infectious bronchitis (IB) vaccine strains (H120, Ma5 and 4/91) ,one reference strain M41 and 26 IB field isolates. These 30 strains were classified in 7 different genotypes, respectively. Virus-neutralizing test on tracheal organ cultures (TOC) with chicken embryo were used to evaluate relatedness values of the antigenicity based on the antibody titer, to analyze the antigenic relationships between the isolates and vaccine strains, as well as to determine the serotypes of 26 IB viruses isolated from the field in Guangxi between 1985 and 2008. The results showed 30 strains were classified into 7 distinct serotypes and there were two predominant serotypes within the 26 isolates, serotypes 1 (totally 13 isolates) and serotype 2 (totally 5 isolates), respectively. In addition, there were some differences observed between the results of serotyping and the genotyping (including the S1, N, M and 3'UTR). The results of the study demonstrated that there were different predominant serotypes and multiple serotypes of IBV circulated in Guangxi in recent years, antigenic variation existed between Guangxi field isolates and vaccine strains.
Animals ; Antibodies, Viral ; immunology ; Antigens, Viral ; genetics ; immunology ; Chick Embryo ; Chickens ; China ; Coronavirus Infections ; immunology ; veterinary ; virology ; Genetic Variation ; Genotype ; Infectious bronchitis virus ; classification ; genetics ; immunology ; isolation & purification ; Phylogeny ; Poultry Diseases ; immunology ; virology ; Viral Envelope Proteins ; genetics ; immunology

The S1 gene hypervariable region I (HVR I) of 22 infectious bronchitis virus (IBV) strains isolated in Guangxi during the period of 1985-2007 were sequenced and compared to that of the other IBV reference strains and the pigeon coronavirus isolates. A phylogenetic tree based on nucleotide sequences of HVR I of all the IBV showed that they were classified into 5 distinct Clusters. 16 out of 22 IBV isolates were grouped into Cluster I, and had higher homology with pigeon coronavirus isolates but lower homology with the Massachusetts (Mass) type vaccine strains. There were 4 and 3 amino-acid residues inserted at the sites of 33-34 and 34-35 respectively within HVR I in 15 isolates, except in isolate GX-NN6 there had 4 amino-acid residues inserted at the both sites; isolates GX-YL1 and GX-NN2 had close relationship with Mass type vaccine strains, and they shared Cluster II; isolates GX-G and GX-XD of Cluster III had close relationship with the Japanese strain JP Miyazaki 89 which was isolated at the same period; isolates GX-YL6 and GX-NN7 of Cluster V had close relationship with the European strain 4/91. The results showed that there were high phylogenetic diversity among the IBVs prevailed in the field in Guangxi resulting from the commonly occurred mutation or insertion within the S1 gene HVR I of the viruses, and majority of the isolates had lower homology with the commonly used Mass type vaccine strains. There was much higher homology among viruses isolated in the same period of time, but without distinct difference in geographical origins.
Amino Acid Sequence ; Animals ; Chickens ; virology ; Genetic Variation ; Infectious bronchitis virus ; classification ; genetics ; isolation & purification ; Membrane Glycoproteins ; chemistry ; genetics ; Molecular Sequence Data ; Phylogeny ; Spike Glycoprotein, Coronavirus ; Viral Envelope Proteins ; chemistry ; genetics

BACKGROUND: This study aimed to investigate the epidemiological and clinical characteristics of patients with heat-related il ness, and guide the prevention, diagnosis and treatment of heat-related il ness. METHODS: From June 2013 to August 2013, seventy patients with heat-related illness were treated at Jinshan Hospital of Fudan University, and their epidemiological characteristics, laboratory results, treatment and prognosis were retrospectively analyzed. RESULTS: In the 70 patients, 18 patients suffered from heat stroke and 52 patients from non-heat stroke. When the environmnent temperature was above 35 °C, the body temperature of the patients began to increase markedly. The patients with heat stroke were significantly older than those with non-heat stroke (P<0.05). The body temperature, heart rate, blood glucose, blood lactate dehydrogenase and blood creatine kinase in the patients with heat stroke were higher than those in the patients with non-heat stroke (P<0.05). Blood lactate dehydrogenase and blood creatine kinase were positively correlated with body temperature (r=0.801). CONCLUSION: When the environmental temperature goes above 35 °C, heat-related illness should be prevented, especially in the elderly. The body temperature, heart rate, blood glucose, blood lactate dehydrogenase and blood creatine kinase in patients with heat stroke are higher than those in patients with non-heat stroke. Blood lactate dehydrogenase and blood creatine kinase are positively correlated with body temperature, but their relationship with heat-related illness awaits further study.

Objective To explore the effect of hydrogen peroxide on the prevention of the infectious complication in the patients with orotracheal intubation.Methods Ninety patients with orotracheal intubation for mechanical ventilation were randomly divided into three groups,each with 30 cases.The oral cavities of the patients in Group A were washed with cotton balls soaked by 3％ hydrogen peroxide solution and rinsed with normal saline (0.9％ sodium chloride solution) ;those of the patients in Group B were rinsed with normal saline alone;and those of the patients in Group C were washed conventionally with cotton balls soaked by normal saline.The oral care was conducted twice daily in each group.The bacterial number in pharyngo-oral cavities,the pH values,clean degrees of oral cavities and the incidence of oral cavity infection in the three groups were observed.Results After oral care with the three methods,the bacterial number in pharyngo-oral cavities of the patients in Groups A,B and C were decreased,and respectively (306.70 ± 15.57),(436.10 ± 19.14),(762.33 ± 28.46) cfu/plate,the difference was statistically significant (F =116.5,P ＜ 0.05),and the decrease in bacterial number was most significant in Group A.The oral pH values of all the patients were increased,and were respectively (6.70 ± 0.085),(6.41 ± 0.102),(6.35 ± 0.076) in three groups,the difference was statistically significant (F=4.415,P＜0.05),and those of the patients in Group A were found to increase most significantly.The incidence of dental plaques (16.67％,40.00％,33.33％) and that of oral cavity infection (10.00％,20.00％,33.33％) did not show statistical significance (x2 =4.127,4.937,respectively; P ＞0.05).The incidence of halitosis (20.00％,43.33 ％,56.67％) showed significant differences (x2 =8.61,P ＜0.05),and the incidence of halitosis was lowest in Group A.Conclusions Washing the oral cavities with cotton balls soaked by 3％ hydrogen peroxide solution and rinsing the oral cavities with 0.9％ sodium chloride solution can prevent from the infectious complication in the patients with orotracheal intubation.

Siraitia grosvenorii is a traditional Chinese medicine also as edible food. This study selected six candidate reference genes by real-time quantitative PCR, the expression stability of the candidate reference genes in the different samples was analyzed by using the software and methods of geNorm, NormFinder, BestKeeper, Delta CT method and RefFinder, reference genes for S. grosvenorii were selected for the first time. The results showed that 18SrRNA expressed most stable in all samples, was the best reference gene in the genetic analysis. The study has a guiding role for the analysis of gene expression using qRT-PCR methods, providing a suitable reference genes to ensure the results in the study on differential expressed gene in synthesis and biological pathways, also other genes of S. grosvenorii.
Cucurbitaceae ; genetics ; RNA, Ribosomal, 18S ; genetics ; Real-Time Polymerase Chain Reaction ; methods ; Reference Standards

Total RNA was isolated from Siraitia grosvenorii fruit by the method of modified Trizol, according to S. grosvenorii fruit characteristics of rich phenols, polysaccharide, oil and proteins. The OD260/280, OD260/230, RNA integrity (RIN) and yield of the total RNA with this method were 2.01, 2.02, 9.50 and 260 mirog.g-1, respectively. The open reading frame (ORF) of dehydroascorbate reductase (DHAR), named as SgDHAR, was cloned by rapid amplification of cDNA ends (RACE) and RT-PCR method from S. grosvenorii. The GenBank accession number for this gene is KC907731. The SgDHAR gene contains a full-length cDNA of 1,252 bp including ORF of 819 bp and encodes a predicted protein of 272 amino acids. The molecular mass is 30.217 7 kD and the isoelectric point is 8.76. Homology comparison showed that it shared 87% nucleotide sequence homology with Cucumis sativus. Expression patterns using qRT-PCR analysis showed that SgDHAR was mainly expressed in fruit and stem, followed by flower, and was lowest in root, while the expression level was 6.83 times in triploid. T than that in diploid. Therefore, SgDHAR gene may be involved in abortion of triploid seedless S. grosvenorii.
Amino Acid Sequence ; Base Sequence ; Cloning, Molecular ; Cucurbitaceae ; chemistry ; genetics ; DNA, Complementary ; genetics ; Flowers ; chemistry ; genetics ; Fruit ; chemistry ; genetics ; Molecular Conformation ; Open Reading Frames ; Oxidoreductases ; genetics ; metabolism ; Phylogeny ; Plant Roots ; chemistry ; genetics ; Plant Stems ; chemistry ; genetics ; Plants, Medicinal ; chemistry ; genetics ; Protein Structure, Secondary ; RNA, Plant

To study the optimal examination method of CD62P and CD63 and investigate platelet activation in patients with diabetes mellitus (DM), whole blood labeled directly with monoclonal antibodies CD62P and CD63 and flow cytometry were used to evaluate the positive percentages and the mean fluorescence intensity of CD62P and CD63. The specimens of peripheral blood obtained from 10 healthy adults were divided into two groups. In the unfixing group, the positive percentages of CD62P and CD63 at the periods of 30, 60, 90 and 120 minutes after staining were (7.57 +/- 2.33)%, (20.50 +/- 5.70)%, (28.70 +/- 5.67)% and (36.52 +/- 6.13)%, and, (0.89 +/- 0.36)%, (1.11 +/- 0.84)%, (2.35 +/- 2.02)% and (5.43 +/- 3.66)% respectively, their respective MFI were 1.57 +/- 0.13, 1.88 +/- 0.08, 2.00 +/- 0.09 and 2.38 +/- 0.22 and 3.91 +/- 0.11, 4.07 +/- 0.16, 4.38 +/- 0.14 and 4.44 +/- 0.19. However, in fixing group with 1% paraformaldehyde, the results had not any obvious change and almost were same. Besides it, the positive percentages of CD62P and CD63 in 37 adult patients with DM were (14.11 +/- 6.68)% and (2.71 +/- 1.74)%, significantly higher than that in the normal controls. It is concluded that the CD62P and CD63 on platelet membrane were very sensitive and would be easily activated in vitro, all manipulations that includes labeling with antibody, incubation and detection using flow cytometry should be finished within 30 minutes after samples collected. While fixing by using 1% paraformaldehyde can steady the labeling compounds and effectively prevent the artificial activation of platelet, and keep the stable results within two hours after the samples labeled. In adult patients with DM, the relationship between the cardiovascular complication of diabetes and platelet activation might be existed.

AIMTo produce poly (lactic-co-glycolic acid) (PLGA) microspheres, containing a staphylokinase variant (K35R, DGR) with reduced immunogenecity and antiplatelet aggregation activities, which allowed the preservation of protein stability during both particle processing and drug release.

METHODSDGR-loaded microspheres were fabricated using a double emulsion-solvent evaporation technique. The effects of preparative parameters, such as stirring rate, polymer concentration, and the excipients of both internal and external aqueous phase (W2), on DGR encapsulation efficiency and microsphere characteristics were investigated. In vitro and in vivo release of DGR were conducted and the cause for instability of DGR during release was also investigated.

RESULTSModerate ultrasonic treatment of aqueous DGR/dichloromethane mixtures caused approximately. Eighty four per cent DGR denaturation. However, the activity recovery of DGR almost amounted to 100% when 2% polyvinyl alcohol (PVA) was addled into the aqueous phase. It was found that NaCl in the external water phase significantly increased DGR encapsulation efficiency. Furthermore, NaCl in the external water phase played a role in determining size and surface morphology of microsphere. In vitro release test showed a burst release of DGR from microspheres, followed by sustained release of 50% total activity over 15 days. In vivo experiments showed that DGR released from microspheres sustained 5 days. Denaturation of DGR within microspheres might be resulted from acidic microclimate.

CONCLUSIONThe stability of DGR was effectively protected during microencapsulation and a relatively high encapsulation efficiency of DGR was obtained. PLGA microspheres could be an effective carrier for DGR.

Animals ; Area Under Curve ; Drug Carriers ; Drug Compounding ; Drug Delivery Systems ; Escherichia coli Proteins ; administration & dosage ; genetics ; pharmacokinetics ; Genetic Variation ; Lactic Acid ; Male ; Metalloendopeptidases ; administration & dosage ; genetics ; pharmacokinetics ; Microspheres ; Particle Size ; Polyglycolic Acid ; Polymers ; Rabbits

Nonaqueous capillary electrophoresis is used for the determination of the contents of diosgenin and ruscogenin in Radix Ophiopogonis. The operating buffer was composed of 20 mmol x L(-1) Na2B4O7-HCl (pH 7.61) in 70% methanol. The applied voltage was 25 kV and detection potential was at +0.70 V. With these conditions, the components were successfully separated. The content of diosgenin in Radix Ophiopogonis was 0.018 mg x g(-1) and ruscogenin was 0.008 mg x g(-1). The average recoveries of diosgenin and ruscogenin were 102% and 99.2%, respectively. A new method of the quality control of diosgenin and ruscogenin in Radix Ophiopogonis is provided.
Diosgenin ; analysis ; Electrophoresis, Capillary ; methods ; Ophiopogon ; chemistry ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Spirostans ; analysis