Objective To investigate the inhibitory effect of siRNA human telomerase transcriptase (hTERT) on activity of telomerase and proliferation of lung carcinoma cell A549. Methods A plasmid including U6 promoter and siRNA of hTERT was designed and constructed. The plasmid was transfected into A549 cell line. The telomerase activity was tested by telomerase repeat amplification protocol ELISA (TRAP-ELISA). MTT assay was used to assay the cell proliferation activity,and hTERT expression was assessed by Western blot. Result The U6 expression plasmid that was constructed for hTERT gene 745 showed obvious interfering effect. hTERT-siRNA could down-regulate the expression of hTERT protein,inhibit telomerase activity and proliferation of A549 cells. Conclusion siRNA of hTERT can inhibit the expression of human telomerase and proliferation of A549 cells. It may open a new approach to the use of siRNA as a new tool to study gene function in cancer cell lines,and may be developed to be a new gene therapeutic agent for cancer.

Nonalcoholic fatty liver disease (NAFLD) is a clinicopathological syndrome caused by liver damage factors except alcohol and has the major feature of diffuse macrovesicular hepatocyte steatosis.Thetwo-hit hypothesis can partly explain the pathogenesis of NAFLD.Recent studies have found that ceramide is a key molecular messenger involved in the development and progression of NAFLD,and as a sphingolipid,it is closely associated with the two-hit hypothesis.This article reviews the role of ceramide in NAFLD.

BACKGROUND:In previous experiments, TiO2-xNy OBJECTIVE:To study the friction properties of orthodontic brackets coated with TiO-coated bracket has been confirmed to have excelent antibacterial properties and biological safety performance. 2-xNy METHODS: TiO film. 2-xNy film was prepared by radiofrequency magnetron sputtering on MBT bracket (0.022″). The TiO 2-x N y RESULTS AND CONCLUSION: TiO-coated brackets were analyzed by X-ray diffraction and scanning electron microscopy observations. The coefficient of static friction and coefficient of kinetic friction between the wires (0.012″, 0.014″, 0.016″) and orthodontic brackets were calculated. 2-xNy film on the bracket was of anatase structure, which was compact and had good crystalinity. Under dry condition, the coefficient of static friction and kinetic friction of the brackets coated with TiO2-xNy were less than those of ordinary brackets; under wet condition, the coefficients of static friction and kinetic friction of the brackets coated with TiO2-xNy were less than those of ordinary brackets, but the difference was not statisticaly significant. Nano-TiO2-xNy film can reduce the friction between bracket and archwire, which wil offer a novel opportunity to significantly reduce the friction during tooth movement.

Applying modern computer network information technology and digital ECG technology,ECG experiment education software system with learning theory,reading ECG picture and practicing report sheet was developed by the Microsoft Visual C++ 6.0 language as the main programming language,and by present widely used relational database MySQL as the backend database.This education software system,used by more than 2 000 different professional students in 2013 and 2014,can be effective to improve their interpretation skills in ECG,and expands the non-class space of ECG education by the modern network technology.It is worthy of popularization as a powerful helper of ECG teaching.

Objective To evaluate the agreement of bispectral index values recorded from tumor side and the contralateral areas during supratentorial craniotomy.Methods Thirty-five patients (16 males, 19 females, aged 18-65 years, ASA physical status Ⅱ or Ⅲ) scheduled for supratentorial tumor resection were enrolled in this study.Bispectral index (BIS) sensors were placed at bilateral frontal areas.The patients were anesthetized with propofol.From each BIS monitor, we collected data at each of four time stages: before the induction of anesthesia, before dura opening, removal of tumor and recovery of conscious.These data were compared using Bland-Altman analysis.Results Bland and Altman analysis revealed a BIS negative-bias (limits of agreement) of before induction-0.8(-7.2-5.7), before dura opening 0.6 (-8.3-9.5), and removal of tumor 1.5(-6.9-9.9), recovery of conscious 0.2 (-9.3-9.8).Conclusion There are significant agreement for BIS values between the frontal area of tumor side and the contralateral areas.BIS values can be used interchangeably between bilateral frontal.

Objective:To study the inhibitory effect of γδT cells on the proliferation of ovarian cancer SKOV3 cells,and to clarify its possible mechanism of inducing apoptosis. Methods:The human ovarian cancer SKOV3 cells cultured in vitro were used as control group,and theγδT and SKOV3 cells were co-cultured for 72 h as γδT cells treatment group.Laser scanning confocal microscope was used to obeserve the morphological changes of nucleus SKOV3 cells,and the inhibitory rate of proliferation of SKOV3 cells in two groups were detected by MTT method;Transwell Chambers was used to detect the cell migration ability,then the apoptotic rates of SKOV3 cells were tested by flow cytometry (FCM).Results:The apoptotic morphology of nucleus of SKOV3 cells in γδT cells treatment group were found under microscope,such as nuclear shrinkage.The MTT resultes displayed that the inhibitory rate of proliferation of SKOV3 cells in γδT cells treatment group was higher than that in control group (P <0.05).The Transwell Chambers results showed that the number of transmembrane cells in γδT cells treatment group was lower than that in control group,and the migration rate was decreased compared with control group (P <0.05).The FCM results showed that the apoptotic rate of SKOV3 cells in γδT cells treatment group was higher than that in control group (P < 0.05 ).Conclusion:γδT cells can inhibit the proliferation and the migration abilities of ovarian cancer SKOV3 cells,and promote the apoptosis.

Objective:To explore the effect of enhanced recovery after surgery(ERAS)-based multidisciplinary collaboration model on shortening the time of forbidden eating before receiving surgery, provide the basis for the selection of the preoperative diet prohibition scheme.Methods:From January 2017 to February 2019, a total of 384 patients who received the operation in Qingdao Municipal Hospital were analyzed retrospectively. The patients who under the traditional preoperative diet prohibition scheme were taken as the control group(156 cases) while those who under the multidisciplinary cooperation mode nursing under the concept of eras were taken as the experimental group(228 cases). The experimental group formulated the perioperative diet prohibition process according to the guidelines of eras, and the experimental group carried out the perioperative diet management for the patients according to the procedure. The difference between the two groups in the time of fasting, hunger, thirst incidence, insulin resistance, temporary stop will be observed and compared.Results:The time of fasting was (4.01±1.55) h in the experimental group and (10.12±1.57) h in the control group,there was significant difference between the two groups( t value was -1.65, P < 0.01). The incidences of thirst,hunger were 13.2%(30/228), 11.8%(27/228) in the experimental group and 89.7%(140/156), 87.2%(136/156) in the control group, there were significant differences between the two groups(χ 2 values were 220.20, 215.20, P < 0.01). The levels of insulin resistance on the first and third day after operation were 1.85 ± 0.43,1.52±0.61 in the experimental group and 1.99±0.51, 1.67±0.49 in the control group, the differences were statistically significant ( t values were -2.90, -2.56, P < 0.05).The temporary stop rate was 1.75%(4/228) in the experimental group and 7.69%(12/156) in the control group, the difference was statistically significant( χ2 value was 8.19, P<0.01). Conclusions:The ERAS-based multidisciplinary collaboration model can effectively shorten the preoperative fasting time, reduce the level of insulin resistance, reduce the incidence of hunger and thirst, and improve the rate of temporary stop and adjustment.

Objective To evaluate the diagnostic value of gray-scale ultrasound combined with contrast-enhanced ultrasound (CEUS) in thyroid imaging and reporting system ( TI-RADS) 3-5 nodules of American College of Radiology ( ACR) . Methods The 208 patients( 216 nodules) who underwent thyroid contrast-enhanced ultrasound examination from April 2014 to December 2016 were retrospectively analyzed . The ACR TI-RADS classification and contrast-enhanced mode of thyroid ultrasound were evaluated . The benign and malignant thyroid lesions were constructed through the analysis of contrast-enhanced mode of thyroid benign and malignant lesions in the past literature and the preliminary study of our group . Prediction model of gray scale combined with contrast-enhanced ultrasound for lesions were established . Besides ,the sensitivity ,specificity and accuracy of gray scale ultrasound combined with contrast-enhanced ultrasound prediction model for differential diagnosis of benign and malignant thyroid lesions were calculated based on the results of puncture biopsy or surgical pathology as the gold standard . Results Compared with pathological results , the positive predictive value , negative predictive value , accuracy , sensitivity and specificity of gray scale combined with contrast-enhanced ultrasound in evaluating thyroid nodules were 90 .8％ ,93 .3％ ,91 .2％ ,98 .8％ and 62 .2％ ,respectively . Conclusions Gray-scale ultrasound combined with contrast-enhanced ultrasound can provide clinical value in differential diagnosis of benign and malignant TI-RADS 3-5 thyroid nodules .

Objective:To investigate the effect of Shentai Tea Polyphenols on memory and its underlying mechanism in healthy elderly adults.Methods:According to the randomized, double blind and placebo-controlled prospective experimental design, 240 healthy middle-aged or elderly subjects with normal objective cognition were recruited in our hospital from April to December 2019 through advertising, and they were randomly divided into an experimental group ( n=120) and a control group ( n=120). Oral administration of Shentai Tea Polyphenols or placebo capsules (lasting for 90 d, 2 times/d, 2 capsules/time) was given to subjects in the two groups. The neuropsychological scale scores and near-infrared brain imaging data of all subjects were collected before and after intervention, and the mean time from baseline data collection before intervention to follow-up data collection after medication was controlled within 6 months. Results:In patients from the experimental group, as compared with those before intervention, scores of delayed recall and recognition memory in the auditory verbal learning test (Huashan version) and Boston naming test, scores of Montreal cognitive assessment scale (basis version) were significantly higher, and scores of Hamilton anxiety scale, shape trails test A and rapid-eye-movement sleep behavior disorder screening questionnaire were significantly lower after intervention ( P<0.05); however, only scores of delayed recall in the auditory verbal learning test (Huashan version) were significantly increased in the control group ( P<0.05). A total of 23 pairs of differences of brain function connection before and after intervention in the experimental group were significantly higher than those in the control group ( P<0.05), mainly involving the function connection of forehead peak network (FPN), and function connection of FPN-default network, function connection of FPN-somatosensory movement network, function connection of FPN-dorsal attention network and function connection of FPN-visual network, as well as the function connection of default network and somatosensory movement network. Conclusion:Shentai Tea Polyphenols can improve cognitive performances including memory, language and executive function, anxiety mood and sleep quality in healthy middle aged or elderly subjects by affecting the functional connections of the networks in the brain.

With the rapid development of gene editing technology, the study of spermatogonial stem cells (SSCs) holds great significance in understanding spermatogenesis and its regulatory mechanism, developing transgenic animals, gene therapy, infertility treatment and protecting rare species. Biogenesis of lysosome-related organelles complex 1 subunit 1 (BLOC1S1) is believed to have anti-brucella potential. Exploring the impack of BLOC1S1 on goat SSCs not only helps investigate the ability of BLOC1S1 to promote SSCs proliferation, but also provides a cytological basis for disease-resistant breeding research. In this study, a BLOC1S1 overexpression vector was constructed by homologous recombination. The BLOC1S1 overexpression cell line of goat spermatogonial stem cells was successfully constructed by lentivirus packaging, transfection and puromycin screening. The overexpression efficiency of BLOC1S1 was found to be 18 times higher using real time quantitative PCR (RT-qPCR). Furthermore, the results from cell growth curve analysis, flow cytometry for cell cycle detection, and 5-ethynyl-2'-deoxyuridine (EdU) staining showed that BLOC1S1 significantly increased the proliferation activity of goat SSCs. The results of RT-qPCR, immunofluorescence staining and Western blotting analyses revealed up-regulation of proliferation-related genes (PCNA, CDK2, CCND1), and EIF2S3Y, a key gene regulating the proliferation of spermatogonial stem cells. These findings strongly suggest that the proliferative ability of goat SSCs can be enhanced through the EIF2S3Y/ERK pathway. In summary, this study successfully created a goat spermatogonial stem cell BLOC1S1 overexpression cell line, which exhibited improved proliferation ability. This research laid the groundwork for exploring the regulatory role of BLOC1S1 in goat spermatogonia and provided a cell platform for further study into the biological function of BLOC1S1. These findings also establish a foundation for breeding BLOC1S1 overexpressing goats.
Animals ; Male ; Goats ; Stem Cells ; Spermatogonia/metabolism* ; Cell Proliferation ; Flow Cytometry ; Testis/metabolism*