This article aimed to study the antigenicity of nucleocapsid proteins (NPs) in six pathogenic phleboviruses and to provide theoretical evidence for the development of serological diagnostic reagents. NPs of six pathogenic phleboviruses were expressed and purified using a prokaryotic expression system and rabbits were immunized with individual recombinant NPs. Cross-reactions among NPs and rabbit sera were determined by both indirect ELISA and Western blotting analyses, and the sera titer was determined by indirect ELISA. Furthermore, sera from SFTS patients were also detected by each recombinant NP as a coating antigen using indirect ELISA. The cross-reactions and the sera titer were subsequently determined. Both the concentration and purity of recombinant NPs of six pathogenic phleboviruses met the standards for immunization and detection. The results of indirect ELISA and Western blotting showed that each anti-phlebovirus NP rabbit immune serum had potential serological cross-reactivity with the other five virus NP antigens. Furthermore, the sera from SFTS patients also had cross-reactivity with the other five NP antigens to a certain extent. Our preliminary study evaluated the antigenicity and immune reactivity of six pathogenic phleboviruses NPs and laid the foundation for the development of diagnostic reagents.
Animals ; Antibodies, Viral ; immunology ; Antigens, Viral ; genetics ; immunology ; Cross Reactions ; Humans ; Nucleocapsid Proteins ; genetics ; immunology ; Phlebotomus Fever ; diagnosis ; immunology ; virology ; Phlebovirus ; classification ; genetics ; immunology ; isolation & purification ; Rabbits

A strain KX6,producing thermotolerant endoglucanase,was isolated from compost. The morpholo-gical identification and 16S rRNA sequence analysis showed it belongs to Streptomyces xylophagus. The production and characterization of endoglucanase from Streptomyces xylophagus KX6 was studied. Maximum endoglucanase yield of 0.538 IU/ml was achieved with medium pH8.0,containing CMC2Na 1.0% as carbon resource,soybean meal 1% as nitrogen resource,2% inoculating volume,30% 250 ml triangle flask bulk for medium volume at 40℃ 200r/min shaker for 48h. The endoglucanase exhibited optimum catalytic activity at pH7.0 and 50℃. The enzyme was stable at 50℃,and able to retain 60% of the full activity,when it was incubated at 60℃ for 1h.The enzyme was stable at pH6.0~7.0. All these findings suggest that the enzyme is a thermotolerant neutral endoglucanase.

Multi-template molecularly imprinted solid phase extraction not only has the advantages of high selectivity, large adsorption capacity, easy preparation, reuse and low environmental pollution, but also can realize the enrichment and separation of many kinds of compounds. It has attracted wide attention in the extraction and separation of traditional Chinese medicine components. This study summarizes the latest development of multi-template molecularly imprinted solid phase extraction. At the same time, based on the classification of active components of traditional Chinese medicine (flavonoids, alkaloids, phenylpropanol, terpenes, etc.), the latest application of multi-template molecular imprinting solid phase extraction in multi-component separation of traditional Chinese medicine was reviewed, with a view to better application of multi-template molecularly imprinted polymer in active multi-component extraction and separation of traditional Chinese medicine and provide reference for the material basic research of the efficacy of traditional Chinese medicine.

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

Animals ; Antineoplastic Agents, Phytogenic ; chemistry ; pharmacology ; Cell Division ; drug effects ; Colchicine ; analogs & derivatives ; chemistry ; pharmacology ; Humans ; Molecular Structure ; Structure-Activity Relationship ; Tubulin ; drug effects ; Tumor Cells, Cultured

OBJECTIVETo evaluate the effect of using cone-beam computed tomography (CBCT) and dental operating microscope (DOM) in treating maxillary molars containing bifurcative canals buccally.

METHODS304 endodontically treated maxillary molars (159 maxillary first molars and 145 maxillary second molars) were included. After preparing access to pulp chamber, the number of canal orifices and location in the pulp chamber floor of each tooth were recorded. For those teeth with bifurcative canals buccally confirmed by preoperative radiographs, the root canals were negotiated by naked eyes firstly, then under DOM according CBCT results. Following working length determination, the root canals were prepared by step-down technique and obturated with cold lateral condensation technique. The efficiency was evaluated with radiographs before, during and after operation.

RESULTSIn 304 maxillary molars, 51 molars were found to have two canal orifices (buccal one and palatal one) in the pulp floor, 30 bifurcative canals buccally (8 upper first molars and 22 upper second molars) were found. CBCT information indicated the level of bifurcation in buccal canals were 3-8 mm under the pulp chamber floors. In 30 maxillary molars, 7 teeth treated by X-rays and eyes could be negotiated, 22 teeth treated by CBCT and DOM could be negotiated and were well instrumented and filled by evaluating with radiographs during and after operation, 8 teeth with deep divergent MB2 canals or calcified canal could not be negotiated.

CONCLUSIONOperative field can be located precisely by CBCT and dental operating microscope that could be effective method in treating these sort of canals.

Cone-Beam Computed Tomography ; Dental Pulp ; Dental Pulp Cavity ; Humans ; Maxilla ; Microscopy ; Molar ; Tooth ; Tooth Root

Adult ; Chyle ; Female ; Filariasis ; complications ; Humans ; Laparoscopy ; Lymphatic System ; surgery ; Male ; Middle Aged ; Retroperitoneal Space ; Urine ; Urologic Surgical Procedures ; methods

Objective To evaluate the effects of teaching mode of “5-stage”cycle combined with teaching pathway on improving clinical practice ability of nursing students and patients′satisfaction.Methods A total of 54 practice nursing undergraduates in Oncology Department from September 201 3 to November 201 3 were divided into observation group (28 cases,teaching mode of “5-stage”cycle combined with teaching pathway)and control group (26 cases,teaching mode of “one-to-one”).Clinical practice ability of nursing students and patients′satisfaction were compared between two groups.Results The comprehensive score of theoretical examination (85.89 ±7.1 9)and examination of practical ability (86.57 ±5.34)of nursing students in observation group were higher than those in control group with a statistically significant difference (P <0.05).The satisfaction of patients in basic nursing,technical operation,health education,service attitude were 78.5%,67.0%,73.5%,63.0%in observation group,which were higher than those in control group (P <0.05).Conclusions The teaching mode of “5-stage”cycle combined with teaching pathway can improve clinical practice ability of nursing students and patients′satisfaction.

OBJECTIVETo provide basis for selecting the suitable method of Down's syndrome biochemical screening in the second trimester pregnancy.

METHODSA total of 30 547 singleton pregnancies between 14 and 20(+ 6) weeks of pregnancy were collected and analyzed for maternal serum alpha-fetoproteins (AFP) and human chorionic gonadotrophin, free beta subunit (beta-HCG) with or without unconjugated estriol (uE3). The screening risks were calculated using the software Lifecycle. The detection rates and the cost of per Down's syndrome detected were calculated and compared. And four different methods were compared in a series of 64 serum samples from Down's syndrome pregnancies.

RESULTS(1) Among the 64 affected cases, the detection rate of Down's syndrome was improved no matter in the double test (DT) or in the triple test (TT) if software Lifecycle (LC) was used to evaluate risks. And it was not suitable to evaluate risks with software 2T-Risks in the triple tests. (2) In the cohort of 30 547 singleton pregnancies, the detection rate of Down's syndrome with project DT-LC, which was double test using AFP and free beta-HCG together with software Lifecycle, and project TT-LC, which was triple test using AFP, free beta-HCG and uE3 together with software Lifecycle, was 56.25% and 57.14%, respectively. The former project was better because it decreased the false positive rate at a lower running cost.

CONCLUSIONThe DT-LC is an effective screening strategy for second trimester detection of fetal Down's syndrome in mainland China.

Adult ; Chorionic Gonadotropin, beta Subunit, Human ; blood ; Down Syndrome ; blood ; diagnosis ; Estriol ; blood ; Female ; Genetic Testing ; methods ; Humans ; Pregnancy ; Pregnancy Trimester, Second ; blood ; Prenatal Diagnosis ; economics ; methods ; Young Adult ; alpha-Fetoproteins ; metabolism

OBJECTIVETo study the chemical constituents of Botrychium lanuginosum.

METHODVarious chromatographic techniques were used to isolate and purify the constituents. The structures were elucidated by chemical evidence and spectroscopic methods.

RESULTTen compounds were isolated from the 95% ethanol extract of the herb of B. lanuginosum and their structures were elucidated as 30-nor-21beta-hopan-22-one (1), beta-sitosterol (2), luteolin (3), thunberginol A (4), apigenin (5), (6'-O-palmitoyl)-sitosterol-3-O-beta-D-glucoside (6), daucosterol (7), 1-O-beta-D-glucopyranosyl-(2S, 3R, 4E, 8Z)-2-[(2R-hydroxy hexadecanoyl) amino]-4, 8-octadecadiene-1, 3-diol (8), luteolin-7-O-glucoside (9), sucrose (10).

CONCLUSIONCompounds 1-10 were isolated from this genus for the first time.

Apigenin ; chemistry ; isolation & purification ; Chromatography ; Drugs, Chinese Herbal ; chemistry ; Ferns ; chemistry ; Isocoumarins ; chemistry ; isolation & purification ; Luteolin ; chemistry ; isolation & purification ; Sitosterols ; chemistry ; isolation & purification ; Sucrose ; chemistry ; isolation & purification