AIM:To study the application of ultrasonic extraction in process of extracting diosgenin from Dioscorea zingiberensis C.H.Wright,and to evaluate its merits.METHODS:A new process of ethanol extracting and hydrochloric acid hydrolysis was provided to extraction diosgenin,and five different extraction methods were studied and compared,and the reaction mechanism was discussed.RESULTS:The ultrasonic extraction method was more efficient.Compared with the traditional water-hydrolylis,extraction rate was 15 percent higher,dosage of hydrochloric acid was registered 90 percent decrease,and the production was purer.CONCLUSION:The grinding pretreatment of Dioscorea zingiberensis is help to enhance the effects of ultrasonic extraction,the method has some advantages in efficiency,energy-saving and low pollution.

Objective:To study the surgical approach to the orbital floor and infraorbital rim.Method:15 cases of fractures of the orbital floor and infraorbital rim were treated with the transconjunctival approach.Result:All fractures of the orbital floor and infraorbital rim were repositioned and fixed through transconjunctival approach. Except for 1 case with mild canthal malposition, no other complication was observed.Conclusion:The transconjunctival approach can provide adequate exposure of the infraorbital rim and orbital floor without scar formation in the face.

Objective: To investigate the changes of urinary S100B protein concentrations and their relationship with brain damage in preterm infants there of . Methods: The urinary S100B protein of 84 preterm infants and 26 full term infants, which were used as control, were measured at 24 h and 120 h after birth. At the same time, routine clinical observations, neurologic patterns and ultrasound screens were recorded. The value of urinary S100B protein and brain damage were evaluated in preterm infants with different gestational age. Results: The differences of urinary S100B protein were statistical significance between the different gestations. The levels of urinary S100B protein were higher in preterm infants, whose gestations were lower than 32 W, than those of other groups. The levels of S100B protein were significantly higher in samples of 27 peri-intraventricular hemorrhage (PIVH) and 3 peri-ventricular leukomalacia(PVL) than those in samples without brain damage( P ＜ 0.05). The S100B levels were significantly higher in urine of 10 preterm infants with polycethemia than those in infants without brain damages. In addition, the S100B levels were different in urine of preterm infants with different prognosis. The S100B levels were significantly higher in urine of infants who died or deteriorated than those of others(P ＜ 0.05). Conclusion: There is an evident trend of decrease in urinary S100B protein concentration with increasing gestational age. It will be helpful to identify preterm infants with PIVH,PVL and high risk of brain damages by measurement of S100B protein in urine early after birth, which indicates further inspection, provides protective treatment and enhances follow up.

Objective To construct and implement theSTARnurse training model, and discuss its application effect and the problems that should be paid attention to, and to provide operational cases and practical basis for nursing clinical education. Methods Through literature review and expert consultation, the framework and content of STAR nurse training model were set up and implemented. The questionnaire survey and semi structured in-depth interviews were conducted among the nurses in the hospital to evaluate the effect of improving the nurses′self-directed learning ability. Results After the implementation of the project, the scores of the three dimensions of self-management, desire for study and self-control were (3.67±0.57), (4.05±0.54), (3.99±0.50) points, which were higher than (3.55±0.49), (3.71± 0.52), (3.53±0.42) points before implementation. The difference was statistically significant (P<0.05 or 0.01). The semi structured in-depth interviews showed that all the nurses believed that STAR nurse training model could promote independent learning and stimulate interest in learning. 14 nurses thought it was beneficial for the nurses to find the problems. Conclusions STAR nurse training model can create a favorable learning environment for nurses, and stimulate the learning motivation. It plays a positive role in improving nurses′ability of self-directed learning.

Objective To construct and implement the nursing quality information feedback system based on QUACERS model and control theory, and discuss its application effect and the problems that should be paid attention to and to provide operational cases and practical basis for nursing quality management. Methods Through literature review and expert consultation, the framework and content of nursing quality information feedback system were set up and implemented. The changes of the nursing quality and the repeated occurrence of nursing problems were evaluated before (2013) and after (2015) the implementation of this project. And in December 2015, a self-made questionnaire was conducted among the nurses in the hospital to evaluate the importance of the feedback and it′s effect of improving the nursing quality. And evaluate the timeliness and effectiveness of different feedback forms. Results 1 120 and 1 136 nurses were followed-up in 2013 and 2015 respectively. The scores of human resource management, clinical nursing service and nursing safety management were higher than before, and the repeated occurrence of nursing problems was lower than before. A total of 450 questionnaires were distributed, 438 copies were collected, the effective recovery rate was 97.33%. Ratings for the importance of each item was from 3.37 to 4.57. Ratings for the effect of improving the quality of care was 3.79 to 4.39. The percentage of quality information received by nurses was more than 95%, and the average score of feedback timeliness was 4.29 to 4.53. Conclusions Quality information feedback system based on QUACERS model can cover multiple dimensions of quality management, and it was conducive to obtaining comprehensive information;Combined with multiple feedback forms can improve the effect of information feedback.

In recent years, Hepatitis B virus (HBV) persistent infection mouse model with recombinant adeno-associated virus 8 carrying 1.3 copies of HBV genome (rAAV8-1.3HBV) is concerned. We studied and compared the efficacy among HBV persistent infection mice models by other serotypes except AAV8. First, we prepared and purified five viruses: rAAV1-1.3HBV, rAAV2-1.3HBV, rAAV5-1.3HBV, rAAV8-1.3HBV and rAAV9-1.3HBV. Then we injected each virus into 3 C57BL/6J mice with the dose of lx 1011 vg (Viral genome, vg) per mouse. We detected HBsAg and HBeAg in sera by enzyme-linked immunosorbent assay (ELISA) at different time points post injection. We killed mice 8 weeks post injection and took blood and livers for assay. We detected copies of HBV DNA by real-time quantitative PCR in sera and livers. Meantime, we detected HBcAg in the livers of mice by immunohistochemistry and further performed pathology analysis of these livers. The five groups of mice, HBeAg and HBsAg expression sustained 8 weeks in serological detection and HBV DNA was both detected in sera and livers at the time of 8 weeks post injection. HBeAg, HBsAg, HBV DNA copies expression levels in descending order were AAV8>AAV9>AAV1>AAV5>AAV2. HBcAg expression was detected in livers as well. Varied degrees of liver damage were shown in five groups of mice. This study provides more alternative AAV vector species to establish a persistent infection with hepatitis B model.
Animals ; Dependovirus ; classification ; Disease Models, Animal ; Enzyme-Linked Immunosorbent Assay ; Genetic Vectors ; Genome, Viral ; Hepatitis B ; virology ; Hepatitis B Core Antigens ; metabolism ; Hepatitis B Surface Antigens ; blood ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; genetics ; Mice ; Mice, Inbred C57BL ; Serogroup ; Virus Replication

Activities of 58 miRNAs for BHK21, HEK293, and Vero cell lines were screened using the high-throughput miRNA activity profiling method. miR-206 activity was found specifically high in BHK21. Considering miR-206 was recognized as a muscle-specific miRNA, we further detected the expression and activity level of miR-206 in BHK21 cells, with myoblast cells C2C12 as positive control and HEK293 cells as negative control. Then, we induced BHK21 by culturing with medium containing 2% horse serum (HS) and tested expression level of slow skeletal myosin heavy chain (MHC), activity and expression levels of miR-206, and expression level of Connexin43 (Cx43) which was reported negatively regulated by miR-206. Results demonstrated that activity and expression levels of miR-206 were both higher in BHK21 cells than in C2C12 cells. After induction of HS, MHC expression level was increased in BHK21 cells. The activity and expression levels of miR-206 were further enhanced. The Cx43 expression level was decreased. These results suggested that BHK21 had the characters of myoblast cells. In conclusion, we firstly discovered that miR-206 activity was specifically high in BHK21 cells, suggesting that BHK21 cells were derived from interstitial cells other than parenchyma cells of kidney from miRNA point of view. Our study also indicated that BHK21 cells were able to be used as models in vitro for research of miR-206 function.
Animals ; Cell Line ; Cercopithecus aethiops ; Connexin 43 ; metabolism ; Cricetinae ; Gene Expression Regulation ; HEK293 Cells ; Humans ; MicroRNAs ; genetics ; Myosin Heavy Chains ; metabolism ; Vero Cells

To overcome the present limitations of passive biochip, based on the basic principle of antigen-antibody reaction, we develop an antigen-antibody reaction model of solid-phase surface and design a novel active biochip system according to this model, which introduces the negative pressure and controlling devices to control the immunoreactions on the nitrocellulose (NC) membrane. From the computer simulation results, this is a rapid, stable, robust and practicable system, which can be used to increase the efficiency of immunoreactions and improve the reproducibility and accuracy of biochip analysis.
Antigen-Antibody Reactions ; Biosensing Techniques ; instrumentation ; Equipment Design ; Models, Biological

To establish an orthotopic transplant mouse model of hepatocellular carcinoma (HCC) labeled with secretary luciferase and to study its response to anti-tumor treatment with interferon-beta gene therapy. We labeled the murine hepatoma Hepal-6 cells with secretary Gaussia princeps luciferase (Gluc), and then injected Gluc labeled Hepal-6 cells intrasplenically in C57BL/6 mice. We monitored blood Glue to evaluate the tumor development and anti-tumor effects of hydrodynamic injection with interferon-beta expressing plasmid. We successfully established the orthotopic mouse model of HCC by intrasplenic injection of Glue labeled Hepal-6 cells. The Glue blood assay could reflect the amount of cancer cells in vivo, tumor progression, as well as anti-tumor effect of interferon-beta gene therapy. In conclusion, Gluc labeled orthotopic transplant mouse model of HCC can ex vivo real-time monitor the tumor development and tumor response to treatments.
Animals ; Carcinoma, Hepatocellular ; pathology ; therapy ; Cell Line, Tumor ; Genes, Reporter ; Genetic Therapy ; Interferon-beta ; genetics ; therapeutic use ; Liver Neoplasms ; pathology ; therapy ; Luciferases ; blood ; genetics ; secretion ; Mice ; Mice, Inbred C57BL ; Neoplasm Transplantation ; Treatment Outcome

We developed a method for monitoring of miRNA activity in live cells by a secreted luciferase gene based plasmid sensor named as Gsensor. Firstly, we constructed pAAV2neo-Gluc-MCS-polyA as "empty Gsensor", which contained multiple cloning sites (MCS) for miRNA target inserted. To detect miR142-3p activity, miR142-3p Gsensor and miR142-3p Gsensor-3 were constructed by inserting one or three complementary miR142-3p targets into pAAV2neo-Gluc-MCS-ployA. Subsequently, miR142-3p Gsensor and miR142-3p Gsensor-3 were respectively transfected into U937 cells and Gluc activity was assayed in the supernatant 48 h post transfection. Results showed that both of them effectively indicated miR142-3p activity of inhibiting Gluc expression compared with empty Gsensor. Simultaneously, miR142-3p Gsensor also demonstrated the inhibition of miR142-3p activity by Anti-miR142 when they were cotransfected into U937 cells. This implied one copy of miRNA target in Gsensor was sensitive enough for investigation of miRNA activity. We further analyzed factors affecting Gsensor function including time and dose, and found that miR142-3p activity sensed by miR142-3p Gsensor rose within 48 h post transfection and approached stable thereafter. Transfected dose varying among 0.001-0.05 pg/cell had little effect on its function. Using miR142-3p Gsensor, we further detected miR142-3p activity in HEK293, U937, K562, SP2/0 and P815 cells. Results suggested that miR142-3p activity was high in U937, K562, SP2/0 and P815 cells and almost negative in HEK293. miR142-3p activity was positively correlated with its relative copies in HEK293, U937 and K562 detected by QRT-PCR. In conclusion, Gsensor proved to be an effective tool for monitoring of miRNA activity in live cells, and provide a new method for monitoring miRNA activity in vitro.
Genes, Reporter ; genetics ; Genetic Vectors ; HEK293 Cells ; Humans ; K562 Cells ; Luciferases ; biosynthesis ; genetics ; MicroRNAs ; metabolism ; Transfection ; U937 Cells