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.

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

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 establish the median of serum markers for second trimester screening in Qingdao region and to assess the influence of median correction on the performance of screening.

METHODSMaternal serum alpha-fetoproteins (AFP), human chorionic gonadotrophin, free beta subunit (β -HCG) and unconjugated oestriol (uE3) were assayed for prenatal screening of 18 188 singleton pregnancies at 15-20(+ 6) weeks gestation from January 2009 to July 2010. The median of serum markers was calculated based on above results and applied for risk estimation in screening for fetal aneuploidy from August 2010 to March 2011. The screening performance, specified in terms of detection rates (DRs), false positive rates (FPRs) and odds of being affected given a positive result (OAPR) were compared between the two groups. The risks of 45 affected pregnancies detected during the study were estimated with both Caucasian and corrected medians.

RESULTSThe average level of AFP in local pregnancies was similar to that of the Caucasian population, whilst β -HCG and uE3 were respectively 11% and 33% higher than those of Caucasians. The multiple of median (MoM) value was between 0.94 and 1.02 for the dataset based on the corrected median. At a cut-off of l in 270, FPR has decreased from 5.2% to 4.9%, and DR of Down syndrome has increased from 60% to 69.2%, and OAPR has increased from 1:79 to 1:59 when evaluating risk based on the corrected median. For the 45 affected pregnancies, three Down syndrome pregnancies could be missed because their risk estimates were lower than the cut-off level based on Caucasian median.

CONCLUSIONIt is useful to establish and apply population and laboratory-specific medians in order to improve the performance of prenatal screening and diagnosis.

Adult ; Biomarkers ; blood ; Estriol ; blood ; Female ; Humans ; Lindane ; blood ; Pregnancy ; Pregnancy Trimester, Second ; Prenatal Diagnosis ; methods ; alpha-Fetoproteins ; analysis

AIMTo investigate the effect of cocaine on apoptosis and caspase-3 activity in germ cells in male rats at different ages.

METHODSCocaine hydrochloride was given (15 mg/kg body weight s.c.) to male Sprague-Dawley rats of 3 weeks (n = 8), 6 weeks (n = 8) and 12 weeks (n = 8) of age, daily for 28 Days. The serum levels of follicle stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL), testosterone (T) and estrogen (E2) were assayed, and the DNA fragmentation of germ cells was determined by gel eletronphoresis. The cell cycle, apoptosis and caspase-3 activity of germ cells were tested by flow cytometry.

RESULTSAfter the 28-day cocaine treatment, testes weight of the 3-week-old rats, the testes and body weights of the 6-week-old rats were decreased significantly compared to those of their corresponding controls (P < 0.05). The serum level of T was decreased significantly in the 3-week-old and 6-week-old rats, and the serum level of PRL was also decreased significantly in 12-week-old rats compared to the controls (P < 0.05). In all the three cocaine-treated groups, the isolated DNA displayed a clear ladder pattern, especially in the 6-week old rats. The number of apoptosic germ cells increased significantly in 3- and 6-week-old rats treated with cocaine (P < 0.05). The caspase-3 activity in all three groups increased significantly compared to the controls (P < 0.05), especially in the 6-week-old rats.

CONCLUSIONCocaine exposure for 28 Days leads to significant damage to male gonad and apoptosis elevation in testes of rats of different ages, especially in those of 6 weeks of age. The increase in caspase-3 activity might be a key pathway related to the early stage of apoptosis as the mechanism of cocaine-induced germ cell loss.

Aging ; physiology ; Animals ; Caspase 3 ; Caspases ; drug effects ; metabolism ; Cell Cycle ; drug effects ; Cocaine ; pharmacology ; Estrogens ; blood ; Follicle Stimulating Hormone ; blood ; Luteinizing Hormone ; blood ; Male ; Prolactin ; blood ; Rats ; Rats, Sprague-Dawley ; Spermatozoa ; cytology ; drug effects ; physiology ; Testis ; drug effects ; pathology ; Testosterone ; blood

OBJECTIVETo investigate the value of magnetic resonance imaging (MRI) in the preoperative assessment of endometrial cancer.

METHODSNinety-four patients with histopathologically confirmed endometrial carcinoma were retrospectively enrolled in this study. MRI findings were compared with the pathologic findings in all cases. The depth of myometrial invasion and lymph node metastasis were evaluated by T2 weighted imaging (T2WI) combined with contrast enhancement or diffusion weighted imaging (DWI).

RESULTSAmong these 94 patients,62 had no or superficial myometrial involvement and 32 cases had deep myometrial involvement. Meanwhile, 24 groups of metastatic lymph nodes and 164 groups of non-metastatic lymph nodes were detected. The accuracy, specificity, sensitivity, negative predictive value, and positive predictive value of T2WI combined with contrast enhancement in discriminating no/superficial myometrial involvement from deep myometrial involvement were 88.3%, 90.3%, 84.4%, 91.8%, and 81.8%, whereas those of T2WI combined with DWI were 81.9%, 87.1%, 71.9%, 85.7%, and 74.2%, respectively. The accuracy, specificity, sensitivity, negative predictive value, and positive predictive value of T2WI combined with contrast enhancement in identifying metastatic lymph nodes were 89.4%, 96.8%, 54.5%, 90.9%, and 78.3%, whereas those of T2WI combined with DWI were 91.5%, 95.5%, 72.7%, 94.3%, and 77.4%, respectively.

CONCLUSIONST2WI combined with contrast enhancement is superior to T2WI combined with DWI in evaluation of the depth of myometrial invasion. However, DWI is more sensitive in identifying lymph node metastasis than T2WI combined with contrast enhancement.

Adult ; Aged ; Endometrial Neoplasms ; diagnosis ; pathology ; Female ; Humans ; Magnetic Resonance Imaging ; methods ; Middle Aged ; Retrospective Studies ; Sensitivity and Specificity