RNA silencing is increasingly employed as an experimental strategy to probe for gene function in several organisms. The purpose of the present study was to test the effect of gene silencing by dsRNA in budding yeast Saccharomyces cerevisiae. GRE3 gene encoding an NADPH-dependent aldose reductase was chosen as an example. A recombinant plasmid psiLENT-GRE3 was constructed based on the pESC-LEU backbone and used to transform S. cerevisiae YPH499. The down regulation of GRE3 gene expression by inducing 1 kb RNA duplex and a 136 bp loop was investigated using reverse transcription- PCR. The results showed that double-stranded RNA mediated gene silencing could be used as a functional tool to decrease the expression level of a specific gene in S. cerevisiae, which would contribute to the understanding of RNA interference in budding yeast.

Proteomics is an emerging discipline developed on the basis of genomics.The fundamental techniques of proteomics include sample preparation,protein separation,protein identification and analysis,and its core techniques are two-dimensional gel electrophoresis and mass spectrometry.In recent years,proteomics has been used in researching the field of Mycobacterium tuberculosis(MTB).Proteomics promotes deep understanding of the pathogenesis of MTB and resistance mechanism via isolating,identifying and analyzing the whole-cell protein and secreted proteins.The development of new vaccine against MTB has showed some promising results based on proteomics.Some powerful early diagnostic markers have been discovered via analyzing the protein composition of MTB clinical isolates.Proteomics also applies to find potential new drug targets,and it has shown many valuable research productions in developing new an-ti-MTB drugs.In summary,the application of proteomics has built a solid foundation for the development of prevention,early diagnosis and treatment of tuberculosis.

Objective To observe the effect of carvedilol on the expression of Cx43 in rabbits with myocardial infarction and its association with ventricular arrhythmia.Method Thirty-six rabbits were randomly divided into three groups in equal number(n=12),namely,myocardial infarction(MI)group,carvedilol group and sham MI group.Rabbits of carvedilol group were administered with carvedilol 5 mg kg~(-1)?d~(-1)after MI induced,while no carvedilol was administered to the MI group.The following observations were made:(1)Cx43 density of the epicardial border zone measured by quantitative immnuoconfocal laser scanning,and(2)cx43 protein expression analyzed by western blot.Results(1)Under immunoconfocal laser microscope,the relative density of Cx43 was(0.16?0.06)% in the infarction group and was(0.32?0.11)% in the sham MI group [(0.68?0.15)%,both P

Objective To develop a new radial artery puncture fixing device to conquer the deficiencies of traditional method. Methods The device was composed of an arm support,a hand support,a wrist part,an arm fixing belt,a palm fixing belt and a tourniquet.The arm support was concave-shaped,and was connected with the hand support with the wrist part.The wrist part had a raised block which was gifted with an inflatable bag.The balloon linked with an inflation balloon with an inflation tube.There was a valve between the inflation tube and balloon.The fixing belt had one end directly connected with the arm support, hand support and wrist part, and the other end linked with the other ends of the above components with Velcro. There was a compression hemostatic balloon at the internal surface of the tourniquet. Results The device behaved well in puncture time and success rate,and decreased the incidence rates of errhysis and hematoma after withdrawing the puncture needle while increased the satisfaction of medical staffs and patients. Conclusion The device gains advantages in simple structure,convenience,practicability and safety,and meets the desired requirements.[Chinese Medical Equipment Jour-nal,2018,39(5):44-46]

Objective To report the surgical method and treatment outecome of transfer of free upper limb lateral bone-skin flap for repair of bone defects and nonunion in hand and forearm.Methods 17 cases of hand composite trauma,5 cases of forearm composite trauma and 2 cases of nonunion in forearm were treated with free bone-skin flaps in distal humerus,whose pedicle was the posterior branch of radical collateral artery. Area of the flap was 2cm?3cm to 8cm?10cm,length of exseeted bone was 3～6 cm.Results All the bone-skin flaps completely survived,the donor area all achieved primary healing,On postoperative 1～4 years follow-up,the texture of the flap was excellent,and bone union was obtained in all transplants,the donor area of distal lateral humerus became thicker and thicker with new cortical bones formed.According to the upper arm function assessment criterion issued by hand surgery association of Chinese medical association.The hand function had excellent results in 17 cases and good results in 2 cases.Conclusion The bone-skin flap has following advantages:easy dissection,reliable blood supply,and no major vessel needed to be sacrificed,so it is an effective method for repair of skin and soft tissue with bone defects in hand and forearm,It.also can be used to repair refractory nonunion in ulna and radius.

Adult ; Female ; HSP27 Heat-Shock Proteins ; HSP70 Heat-Shock Proteins ; biosynthesis ; blood ; HSP90 Heat-Shock Proteins ; biosynthesis ; blood ; Heat-Shock Proteins ; biosynthesis ; blood ; Hepatitis B Core Antigens ; blood ; Hepatitis B, Chronic ; metabolism ; pathology ; Humans ; Liver ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Proteins ; biosynthesis ; blood

OBJECTIVETo understand the levels of total physical activities among different populations, and the distribution of four domains.

METHODSWith proportional stratified sampling and cluster sampling method, 3555 subjects were selected including officials, company staff, high school students, community population and floating population.

RESULTSThe proportions for inactive, insufficiently active, minimally active and health enhancing physical active (HEPA active) as a whole were 20.5%, 10.1%, 26.5% and 42.9% respectively. Community population had the highest level of HEPA active which was 65.6%. Floating population had the highest level of inactivity of 33.1%. Apart from the male floating population, the active level in transportation, physical activity among the investigated were the highest compared to other three active domains. Leisure-time physical activity was in the opposite.

CONCLUSIONThe type of physical activities among general citizens were mainly of physical activity related to transportation but less leisure-time physical activity was seen. Floating population had the highest level of both HEPA activity and inactivity.

Administrative Personnel ; China ; Exercise ; Female ; Humans ; Leisure Activities ; Life Style ; Male ; Students ; Transportation

OBJECTIVETo study the chemical constituents from the whole plant of Aster poliothamnus.

METHODSeparating the chemical constituents by means of chromatography and identifying ther structures on basis of chemical and spectral tecnology.

RESULT AND CONCLUSIONA new triterpene saponin was isolated and identified.

Aster Plant ; chemistry ; Molecular Structure ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Saponins ; chemistry ; isolation & purification

OBJECTIVETo investigate the association between pulmonary vascular remodeling and expression of hypoxia-inducible factor-1α (HIF-1α), endothelin-1 (ET-1) and inducible nitric oxide synthase (iNOS) in pulmonary vessels in neonatal rats with hypoxic pulmonary hypertension (HPH).

METHODSA neonatal rat model of HPH was established as an HPH group, and normal neonatal rats were enrolled as a control group. The mean pulmonary arterial pressure (mPAP) was measured. The percentage of medial thickness to outer diameter of the small pulmonary arteries (MT%) and the percentage of medial cross-section area to total cross-section area of the pulmonary small arteries (MA%) were measured as the indicators for pulmonary vascular remodeling. The immunohistochemical reaction intensities for HIF-1α, ET-1 and iNOS and their mRNA expression in lung tissues of neonatal rats were measured. Correlation analysis was performed to determine the relationship between pulmonary vascular remodeling and mRNA expression of HIF-1α, ET-1 and iNOS.

RESULTSThe mPAP of the HPH group kept increasing on days 3, 5, 7, 10, 14, and 21 of hypoxia, with a significant difference compared with the control group (P<0.05). The HPH group had significantly higher MT% and MA% than the control group from day 7 of hypoxia (P<0.05). HIF-1α protein expression increased significantly on days 3, 5, 7 and 10 days of hypoxia, and HIF-1α mRNA expression increased significantly on days 3, 5 and 7 days of hypoxia in the HPH group compared with the control group (P<0.05). ET-1 protein expression increased significantly on days 3, 5 and 7 days of hypoxia and ET-1 mRNA expression increased significantly on day 3 of hypoxia in the HPH group compared with the control group (P<0.05). Both iNOS protein and mRNA expression were significantly higher on days 3, 5 and 7 days of hypoxia than the control group (P<0.05). Both MT% and MA% were positively correlated with HIF-1α mRNA expression (r=0.835 and 0.850 respectively; P<0.05).

CONCLUSIONSPulmonary vascular remodeling is developed on day 7 of hypoxia in neonatal rats. HIF-1α, ET-1 and iNOS are all involved in the occurrence and development of HPH in neonatal rats.

Animals ; Animals, Newborn ; Endothelin-1 ; analysis ; physiology ; Hypertension, Pulmonary ; etiology ; metabolism ; pathology ; Hypoxia ; complications ; Hypoxia-Inducible Factor 1, alpha Subunit ; analysis ; physiology ; Immunohistochemistry ; Nitric Oxide Synthase Type II ; analysis ; physiology ; Pulmonary Artery ; chemistry ; pathology ; Rats ; Rats, Wistar

Objective To establish a rapid assay with high sensitivity and specificity based on the sequences for group specific gene (GS) and pathogenicity island pag A gene.Methods The PCR primers and probes were designed after the whole sequence was systemically analyzed with bio-informafion tools and blasted with Genebank database.The amplicons were inserted into plasmids so that they could be used as the standard templates to evaluate the sensitivity of the diagnostic system.This assay was based on TaqMan probes and portable Smartcycle PCR machine.Results The detection level was approximately 100 copies per reaction.There was no cross-reaction with other species of Bacillus.This assay could be completed in one hour in laboratory.Conclusion The duplex TaqMan PCR assay could be used to detect Bacillus anthracis rapidly with high sensitivity and specificity.