The Telomeric Repeat Amplification Protocol(TRAP)and its modified versions change the size and/or the ratio of the telomerase products in the amplification stage of the assay.Based on TRAP,a useful method was developed for detecting telomerase activity in rice.A special precursor primer and a special reverse primer and conducted two steps of PCR cycles were designed.GENE Genius? Bio-imaging System was applied for this quantitative analysis for exploring telomerase activity and its optimal reaction conditions.The method ensured that the optimal reaction conditions for the telomerase was 19℃,13minutes,at a concentration of 0.28 u g/? l.A quantitative analysis method was established for detecting telomerase activity in rice.With this method,we detected telomerase activity in roots,young leaves and young panicles of six parental lines of hybrid rice.The results show that young panicles have the highest telomerase activity,demonstrating that telomerase activity is closely related to the cell vitality in plants.

To optimize the water extraction process for glycyrrhiza. Methods: HPLC was used to determine the con-tents of glycyrrhizic acid and liquiritin. The comprehensive index included the contents of glycyrrhizic acid and liquiritin and the yield of dry extract. The water amount and the extraction time were selected as the independent variables, and the comprehensive index was set as the dependent variable. Design-expert 8. 06 software was used to fit multivariate linear or quadratic multinomial models for the experimental values. Response surface methodology was used to optimize the water extraction process. The prediction was carried out through comparing the observed and predicted values. Results:The regression coefficient of binomial fitting complex model was as high as 0. 979 7. The optimum conditions of extraction process were as follows:12-fold amount of water, extracting 3 times with 90 min for each time. The deviation between the observed and predicted values was -1. 72%. Conclusion: Central composite design-response surface methodology is convenient and highly predictive in optimizing the water extraction process for glycyrrhiza, which can be applied in the further membrane separation and purification.

Objective:To explore the effect of different separation and purification technology on the physical and chemical proper-ties of water extract of ophiopogonin. Methods:The water extraction process of total ophiopogon saponins was optimized by an orthogo-nal test. The macroporous resin adsorption and membrane separation technology were adopted to purify the ophiopogonin water extract. The physical and chemical parameters, such as electrical conductivity, pH value, viscosity and turbidity, and the contents of total sap-onins, proteins, tannins and polysaccharides were determined. Results:The optimum water extraction technology of total saponins was as follows:6-fold amount of water was added, boiling 3 times with 90 min for each time. The electrical conductivity, pH value and vis-cosity of different purified liquid of total saponins had no significant differences, while the contents of total saponins and the three kinds of macromolecules showed significant differences. Conclusion: The content of macromolecules can be used as the reference index of purification process of total saponins water extract. Compared with macroporous resin separation, membrane separation technology is more suitable for the separation and purification of total saponins water extract.

To obtain human antibodies against the Gn protein of Severe fever with thrombocytopenia syndrome virus (SFTSV) with phage display technology, this study aimed to screen anti-Gn protein antibodies from an anti-SFTSV Fab human phage display library. Antibody genes were identified by sequence analysis and the specificity of antibodies was confirmed by ELISA. The Fab antibody genes were cloned into the HL51-14 vector and expressed in a mammalian cell expression system. IgG antibodies were then purified by protein A affinity chromatography,and the results were further confirmed by ELISA,IFA,western blotting assays and micro-neutralization tests. The results showed that, after three rounds of panning, there were 390 human Fab antibodies against SFTSV particles, of which 364 were specific for nucleoprotein. Coated with the Gn protein, eight different Fab antibodies specific for Gn protein were obtained after the determination of the subtype and subclass of antibodies by gene sequencing; five of these antibodies were from the Lambda library and three were from the Kappa library. The eight IgG antibodies could specifically bind to Gn protein according to the ELISA, IFA and Western blotting assays. The micro-neutralization test showed that these eight antibodies had no neutralizing activity,but they could still provide a reference for research in human monoclonal antibodies against SFTSV.
Antibodies ; genetics ; immunology ; Bunyaviridae Infections ; genetics ; immunology ; virology ; Cell Line ; Cloning, Molecular ; Humans ; Immunoglobulin Fab Fragments ; genetics ; immunology ; Immunoglobulin G ; genetics ; immunology ; Neutralization Tests ; Phlebovirus ; genetics ; immunology ; Viral Proteins ; genetics ; immunology

Otolaryngology ; Periodicals as Topic ; Quality of Life ; Sickness Impact Profile

Pinellia ternate and Pinellia pedatisecta lectins with biological functions could be obtained through procaryon expression.Their biological similarities and differences were analyzed and further experiments were conducted to discuss the agglutinative activity difference between polymer lectin and metamer lectin.The results showed that agglutinative activity of Pinellia pedatisecta was four times more than that of Pinellia ternate.The differences of the agglutinative activity and pharmacologic action were mainly because two site mutations of Pinellia pedatisecta in the third active site compared with the amino acids sequence in Pinellia ternate.In addition,the metamer lectin expressed by prokaryotic expression system did not agglutinate rabbit red blood cells.It conduct further discussion about the difference between two lectins and the difference between polymer lectin and metamer lectin.It would be applicable for solving the problem of lacking Pinellia resources.

Objective:To investigate the therapeutic effect of carbon dioxide laser tonsillectomy.Method:In this prospective,randomized study, One hundred and two patients were divided into laser group or control group. Patients of laser group were cured with carbon dioxide laser tonsillectom,and the control group was cured with routine method. All operations are executed by one person. Observation index included operation time, hemorrhage in operation, ache after operation, inflammatory reaction of raw surface, repair time of raw surface, rehaemorrhagia and scar.Result:Laser group had advantages of less operation time, less hemorrhage, less ache and less inflammatory reaction of raw surface. Laser group have hemorrhage in operation (7.2±2.1)ml, while control group have hemorrhage in operation (92.0±35.0)ml. Laser group have pseudomembrane early but desquamate late.Conclusion:Carbon dioxide laser tonsillectomy is effective to relieve pain, inflammatory reaction and with less time ,it's an safe , efficient and mini-trauma operation.

This study was aimed to observe renoprotective effect and possible mechanism on Y i-Shui Sheng-Xin Yin in spontaneously hypertensive rats. Sixty 12-week male SHR rats were randomly divided into six groups , which were the Y i-Shui She ng-X in Y in low-dose group , middle-dose group , high-dose group , Benazepril group , model group and blank control group , and ten rats for each group . The SHR rats were sacrificed after eight weeks . The urine microalbumin , blood urea nitrogen and cystatin were tested in each rat . The HE and Masson staining method were used to observe changes of renal pathology . Changes of expression of transforming growth factor-β1 ( TGF-β1 ) , connective tissue growth factor ( CTGF ) , FN were detected by immunohistochemistry . The results showed that compared with the blank control group , blood pressure in model group was associated with a significant rise after 8 weeks. Compared with the model group, blood pressure in the Yi-Shui Sheng-Xin Yin middle-dose group, high-dose group and Benazepril group significantly decreased. Compared with the blank control group , urine microalbumin , blood urea nitrogen and cystatin in model group were associated with a significant rise . Compared with the model group , urine microalbumin , blood urea nitrogen and cystatin in the Y i-Shui She ng-X in Y in middle-dose group , high-dose group and Benazepril group significantly decreased . Pathological examinations showed that pathological changes in model group were faster than all drug-groups , appeared pathological changes of glomerular hypertrophy , glomerular basement membrane thickening of heterogeneity and extensive vacuoles degeneration . Immunohistochemical staining showed that compared with the blank control group , expressions of TGF-β1 , CTGF and FN of rat kidney tissue in model group were obviously up-regulated ( P < 0 . 05 ) . Compared with the model group , expressions of TGF-β1 , CTGF and FN in the Y i-Shui She ng-X in Y in , middle-dose group , high-dose group and Benazepril group were down-regulated ( P < 0 . 05 ) . It was concluded that Y i-Shui She ng-X in Y in can reduce SHR rats' early renal glomerulosclerosis and renal interstitial fibrosis , which play roles of delaying the progress of hypertension and protecting kidney . Its mechanism of action may be related to TGF-β1 , CTGF , FN signal pathways .

Adolescent ; Adult ; Aged ; Endoscopy ; Female ; Humans ; Male ; Maxillary Sinus ; surgery ; Middle Aged ; Otorhinolaryngologic Surgical Procedures ; methods ; Young Adult

To understand the immunogenicity of purified inactivated severe fever with thrombocytopenia syndrome bunyavirus (SFTSV), concentration by ultrafiltration as well as molecular-sieve chromatography (MSC) were used for purification of inactivated SFTSVs. Inactivated viruses in purified samples were analyzed and identified by western blotting and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), the glycoprotein (GP) and nucleoprotein (NP) antigen titers of which were detected using a double-sandwich enzyme-linked immunosorbent assay (ELISA). Purified inactivated SFTSVs were enriched and observed by electron microscopy, and the total protein concentration detected using the bicinchoninic acid assay. Purified inactivated SFTSVs were applied to New Zealand rabbits via two immunization programs to evaluate immunogenicity and to compare the immune effect. After SFTSVs were inactivated and concentrated by ultrafiltration, MSC revealed two typical elution peaks. The sample of one peak was identified as inactivated virions, in which GP and NP were detected by SDS-PAGE, western blotting and ELISA. Main corponent of the other peak was NP. After concentration by ultrafiltration, purified inactivated SFTSVs with purity >90% and total protein concentration of 1. 1 mg/mL were obtained, and the typical electron microscopy of bunyavirus was observed. In the sera of animals immunized with purified inactivated SFTSVs, SFTSV-specific IgG antibody and neutralizing antibody were detected at high titers. However, antibody titers were affected by the immunization program. Effect of immunization on days 0, 14 and 28 was significantly better than that on days 0, 7 and 28. Our work revealed that cultivation of SFTSVs contained intact virus particles and large amounts of free NP. Using MSC, purified inactivated SFTSVs of high purity could be obtained. Purified inactivated SFTSVs induced high titers of neutralizing antibody and virus-specific IgG antibody showing satisfactory immunogenicity, which provides important clues for further study on a vaccine for the inactivated virus.
Animals ; Antibodies, Neutralizing ; immunology ; Antibodies, Viral ; immunology ; Bunyaviridae Infections ; immunology ; virology ; Humans ; Neutralization Tests ; Phlebovirus ; classification ; genetics ; immunology ; isolation & purification ; Rabbits