Objective To study the quality difference of various processing methods about Angelica dahurica. Methods Samples were selected from different places, clean silt by washing and brushing, cut slices and cut blocks in equal division, dried and comminution, to determine total ash, ethanol thermal extract, imperatorin content and HPLC fingerprint similarity. Results The content of total ash was the lowest in “washed”and “washed & cut slice” sample, dilute ethanol thermal extract was the highest in“cut blocks”and“washed&cut blocks”sample, imperatorin content was the highest in“non-washed”and“non-washed & cut slice” sample. Conclusion Washed and cut process is not suitable in place of origin about Angelica dahurica.

To evaluate the adjuvant effect of recombinant enterovirus 71 (EV71) subunit vaccine formulated with chitosan, rabbits were orally immunized with recombinant VP1 (rVP1) or rVP1 mixed with chitosan adjuvant. Levels of virus-specific IgG and IgA antibodies in sera, mucosal wash buffer (intestine, nasal cavity, and lung), and feces were determined by indirect enzyme-linked immunosorbent assay (ELISA). The titers of neutralizing antibodies against EV71 were determined using cytopathic effect-based neutralizing assay, and levels of cytokines (IFN-gamma and IL-4) secreted from in vitro-cultured rabbit splenic lymphocytes under antigen stimulation were also determined by ELISA. Results showed that immunization with rVP1 alone could only induce low levels of serum IgG and mucosal IgA, while rVP1 combined with chitosan adjuvant were able to induce significantly higher levels of antibodies, rVP1 can only induce neutralizing antibodies when used in combination with chitosan. Levels of IFN-gamma and IL-4 in the group immunized with rVP1 plus chitosan were significantly higher than those in the group immunized with rVP1 only or those in the control groups. Our study lays the foundation for development of oral VP1 vaccine against EV71 infection.
Adjuvants, Immunologic ; administration & dosage ; Animals ; Antibodies, Viral ; immunology ; Chitosan ; administration & dosage ; immunology ; Enterovirus A, Human ; genetics ; immunology ; Enterovirus Infections ; immunology ; prevention & control ; virology ; Female ; Humans ; Rabbits ; Vaccination ; Vaccines, Subunit ; administration & dosage ; genetics ; immunology ; Viral Proteins ; administration & dosage ; genetics ; immunology ; Viral Vaccines ; administration & dosage ; genetics ; immunology

OBJECTIVETo study the subcellular localization of severe fever with thrombocytopenia syndrome virus (SFTSV) in macrophages and understand the replication and assembly mechanism of SFTSV in host cells.

METHODSUsing two types of human macrophage cell lines THP-1 and U937, the study analyzed the intracellular colocalization of SFTSV with Golgi apparatus and endoplasmic reticulum by immunefluorescence staining and confocal microscopy.

RESULTSSFTSV infected macrophage cell lines THP-1 and U937. Immunofluorescence staining showed that the SFTSV nuclear protein colocalized with Golgi apparatus and closely surrounded by endoplasmic reticulum in the perinuclear region.

CONCLUSIONThe results suggested that Golgi complex and endoplasmic reticulum are probably the sites for formation and maturation of SFTSV viral particles.

Bunyaviridae ; isolation & purification ; Cell Line, Tumor ; Endoplasmic Reticulum ; virology ; Fever ; virology ; Golgi Apparatus ; virology ; Humans ; Macrophages ; virology ; Thrombocytopenia ; virology

OBJECTIVETo study the mechanism of plant growing promoted by Mycorrhizal fungus through the difference of proteomes.

METHODThe differential proteomes between uninoculated and inoculated endophytic fungi, Epulorhiza sp. on Anoectochilus roxburghii were analyzed by two-dimensional gel electrophoresis and MALDI-TOF/TOF mass spectrum.

RESULT AND CONCLUSIONTwenty-seven protein spots were analyzed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Twenty-two candidate proteins were identified by database comparisons. The function of these proteins mostly involved in signal transduction, metabolic regulation, as well as photosynthesis and substance metabolism. The results indicate that the regulator control system of plant is influenced by fungi action, and the positive regulation improves substance metabolism and photosynthesis, which results in strong plant and higher resistance. It is also deduced that silent genes may exist in endosymbiosis plants.

Basidiomycota ; physiology ; Electrophoresis, Gel, Two-Dimensional ; Host-Pathogen Interactions ; Mycorrhizae ; physiology ; Orchidaceae ; growth & development ; metabolism ; microbiology ; Plant Proteins ; analysis ; Proteome ; analysis ; Proteomics ; methods ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

OBJECTIVETo obtain recombinant human anti-EV71 antibodies from a EV71-associated hand-foot-and-mouth disease patient-derived antibody phage library.

METHODSA combinatorial human scFv library to enterovirus 71 (EV71) virus was constructed using antibody genes harvested from the blood of EV71 virus patients. The library was panned and selected by using purified VP1 protein of EV71 virus with phage display. After that the specific antibody was converted to full human IgG antibody with recombinant baculovirus/insect cell system.

RESULTSOne unique human scFv antibody specific for EV71 virus VP1 protein was obtained by ELISA, IFA and analysis of the antibody DNA sequence. The specific anti-VP1 human scFv antibody was converted to full human IgG antibody with recombinant baculovirus/insect cell system. The full human IgG antibody was tested in vitro for EV71 virus neutralization, resulting in no neutralizing activity with EV71 A type and EV71 C4 subtype.

CONCLUSIONThe obtained human anti-EV71 antibodies without neutralizing activity laid the foundation for diagnosis of human EV71-associated hand-foot-and-mouth disease.

Antibodies, Viral ; immunology ; Enterovirus ; immunology ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunoglobulin G ; immunology ; Peptide Library

OBJECTIVETo develop an assay for titration of severe fever with thrombocytopenia syndrome virus (SFTSV) based on double antibody sandwich ELISA.

METHODSA double antibody sandwich ELISA was developed for detection of SFTSV based on SFTSV nucleocapsid (N) protein specific poly- and monoclonal antibodies, procedures were optimized and evaluated. This ELISA based titration assay was compared with fluorescence assasy and plaque assay based titration method.

RESULTSThe results suggested that the titers obtained by ELISA based method are consistent with those obtained by IFA based method (R = 0.999) and the plaque assay titration method (R = 0.949).

CONCLUSIONThe novel ELISA based titration method with high sensitivity and specificity is easy to manage and perform, and can overcome the subjectivity associated with result determination of the fluorescence assay and plaque assay based methods. The novel ELISA based titration method can also be applied to high throughput detection.

Bunyaviridae ; isolation & purification ; Enzyme-Linked Immunosorbent Assay ; methods ; Fever ; virology ; Fluorescent Antibody Technique ; Humans ; Thrombocytopenia ; virology

Severe fever with thrombocytopenia syndrome bunyavirus (SFTSV) is a novel phlebovirus, causing a life-threatening illness associated with the symptoms of severe fever and thrombocytopenia syndrome. The sequence and structure of the genome have already been illustrated in previous study. However, the characteristics and function of the structure and non-structure proteins is still unclear. In this study, we identified the density of the purified SFTSV virions as 1.135 g/mL in sucrose solution. Using RT-PCR method, we amplified the full coding sequence of RNA dependent RNA polymerase(RdRp), glycoprotein precursor (M), glycoprotein n (Gn), glycoprotein c (Gc), nuclear protein (NP) and non structural protein (NSs) of SFTSV (strain HB29). Respectively inserted the target genes into eukaryotic expression vector pcDNA5/FRT or VR1012, the target protein in 293T cell were successfully expressed. By analyzing the SFTSV virions in SDS-PAGE and using recombinant viral proteins with SFTS patients sera in Western blotting and Immunofluorescent assay, the molecule weight of structure and non-structure proteins of SFTSV were defined. The study provides the first step to understand the molecular characteristics of SFTSV.
Bunyaviridae Infections ; virology ; Cell Line, Transformed ; Fever ; virology ; HEK293 Cells ; Humans ; Orthobunyavirus ; genetics ; metabolism ; Thrombocytopenia ; virology ; Viral Nonstructural Proteins ; biosynthesis ; genetics ; Viral Structural Proteins ; biosynthesis ; genetics ; Virion ; genetics ; metabolism

Objective To investigate the risk factors of aggravated cerebral edema after meningioma surgery.MethodsRespectively analyze the clinical data of 187 patients received neurosurgery operation in our hospital from January 1, 2016 to February 5, 2018 and their postoperative aggravated cerebral edema, the related risk factors for brain edema after meningioma surgery was summarized.Results The incidence of aggravated cerebral edema in patients without preoperative edema (26.23％) was higher than that in patients with preoperative edema (13.8％), the difference was not statistically significant, probably due to the small number of cases or other related factors.Multivariate analysis of all related factors found that preoperative edema was the influencing factor for the increase of brain edema after meningioma surgery (P=0.005).It was found by single factor analysis that tumor site was a risk factor for the aggravation of cerebral edema after meningioma surgery.Multivariate analysis and multiple rate comparisons revealed that the sagittal sinus falx area was an independent risk factor for the aggravation of cerebral edema after meningioma surgery.ConclusionThe presence of peritumoral edema before surgery may be a protective factor for the postoperative brain edema.The incidence of postoperative cerebral edema was significantly higher in meningiomas located near the sagittal sinus falx than that of other sites.Therefore, meningiomas located near the sagittal sinus falx should be attached great importance.During the operation, the venous drainage should be protected, the perioperative management should be strengthened, and hormone and subsequent dehydration should be given timely to improve the prognosis of patients.

OBJECTIVETo investigate the effects of platelet-rich plasma (PRP) on the proliferation of dermal papilla cells (DPCs) and hair follicle regeneration.

METHODSPRP was prepared using the double-spin method and applied to DPCs. The proliferative effect of activated PRP on DPCs was measured using MTT assay. To understand the influence of activated PRP on the hair-inductive capacity of DPCs, freshly isolated epidermal cells and DPCs of passage 4 were resuspended, mixed with various concentrations of a PRP (0%, 5% or 10%) and were then transferred to a grafting chamber, which was implanted onto the dorsal skin of nude mice. The chambers were removed 1 week after grafting and HF formation was monitored for 4 weeks; the graft site was harvested and processed for histological examination.

RESULTSActivated PRP increased the proliferation benefited the aggregative growth of DPCs. There are significant difference in the yield of hair follicles compared with 10% PRP (344 +/- 27) with 0% PRP (288 +/- 35) in the area of reconstituted skin (P < 0.05). The areas treated with PRP demonstrated an increase in hair follicles density of 19.4%. Ten percent PRP (18 +/- 1) d also can significantly shorten the time of hair formation, compared with 0% PRP (20 +/- 1) d (P < 0.05).

CONCLUSIONSThere is a considerable effect of PRP on the time of hair formation and the yield of hair follicles reconstitution.

Animals ; Cell Proliferation ; Cells, Cultured ; Female ; Hair Follicle ; cytology ; growth & development ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Mice, Nude ; Platelet-Rich Plasma ; Regeneration ; Skin ; cytology ; Skin, Artificial

OBJECTIVETo explore the relationship between methyl-tetra-hydrofolic acid (MTHFR) 677 gene polymorphism and the risk of stomach cancer.

METHODSA population based case-control study was conducted and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was used to detect its genotypes.

RESULTSAmong cases with stomach cancer, the frequency of C/C, C/T, T/T genotype were 25.8%, 54.6%, 19.6%, compared with controls as 34.5%, 50.9%, 14.6% respectively. Using C/C genotype as reference, the OR of C/T or T/T genotype was 1.52 (95% CI: 1.04 - 2.23). 53.3% C and 46.7% T allele were distributed in stomach cancer cases, while 60.0% C and 40.0% T in controls. The OR for T allele in relation to C allele was 1.31 (1.02 - 1.69) when C allele was used as reference. In addition, the present study showed that MTHFR677 AnyT genotype might interact with smoking, moldy food intake, wheat porridge intake, eating salty food and Hp CagA infection to increase the risk of stomach cancer. No interaction was observed between MTHFR677 AnyT genotype and alcohol drinking or green tea intake.

CONCLUSIONMTHFR677 AnyT genotype, might increase the risk of stomach cancer development and the genotype might also interact with other environmental risk factors to increase the risk of stomach cancer.

Adult ; Alleles ; Case-Control Studies ; China ; epidemiology ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Life Style ; Male ; Methylenetetrahydrofolate Reductase (NADPH2) ; genetics ; Point Mutation ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Risk Factors ; Smoking ; adverse effects ; Stomach Neoplasms ; enzymology ; etiology ; genetics