The aim of this study is to clone the mouse T-bet promoter and enhancer, construct and identify the firefly luciferase reporter gene plasmid pGL4.10-TBX21pr-CNS for T-bet transcription regulation study and its function in signaling of multiple sclerosis. The promoter and CNS of T-bet were predicted by bioinformatics assay. The predicted fragment of mouse T-bet promoter plus CNS was amplified by PCR and cloned into pGL4.10. The recombinant plasmid pGL4.10-TBX21pr-CNS was transferred into Escherichia coli DH5α. The positive clone was identified by double digestion with Kpn I and Sfi I and DNA sequencing. Finally, pGL4.10-TBX21pr-CNS was cotransfected with pRL-TK into 293T cells and Jurkat cells, pRL-TK and pGL4.10 as a control. The luciferase activity in 293T cells (P = 0.012 2) and Jurkat cells (P = 0.002 2) was higher than that of the control group. A fragment of 1 028 bp mouse T-bet promoter plus 1 308 bp CNS was successfully cloned and the firefly luciferase reporter gene plasmid pGL4.10-TBX21pr-CNS was constructed. In 293T cells and Jurkat cells, pGL4.10-TBX21pr-CNS has the promoter functions. This work offers a basic material for the research of T-bet transcription.
Animals ; Gene Expression Regulation ; Genes, Reporter ; Genetic Vectors ; Luciferases ; Mice ; Multiple Sclerosis ; Plasmids ; Promoter Regions, Genetic ; T-Box Domain Proteins ; genetics

OBJECTIVE:To establish a HPLC method for the determination of chlorogenic acid and baicalin in xiaoer qingrening granula.METHODS:The determination was performed on Symmetry Shield RP C 18 ,the mobile phase of chlorogenic acid consisted of acetonitrile-0.4%phosphoric acid soluton(13∶87)and that of baicalin consisted of methanol-water-glacial acetic acid(50∶50∶1),the detection wavelengths of chlorogenic acid and baicalin were327nm and274nm,respectively,the flow rate was1.0ml/min,sample size was20?l and column temperature was25℃.RESULTS:The respective linear sample size range for chlorogenic acid and baicalin was0.1040?g～1.040?g(r=0.9998)and0.6240?g～6.240?g(r=0.9997),with average recoveries respectively at96.4%(RSD=1.4%)and98.0%(RSD=1.1%).CONCLUSIONS:The method is simple,accurate,reproducible and specific,and it can be used for the quality control of xiaoer qingrening granula.

OBJECTIVE: To investigate the perioperative treatments of endoscopic sinus surgery for nasal diseases in patients with coronary heart disease after PCI. METHOD: Clinical data of 12 cases of endoscopie-assised surgery such as nasal tumors resection,functional sinus surgery,correction of deviated nasal septum,low-temperature plasma hemostasis in patients with coronary heart disease after PCI were analyzed retrospectively. RESULT: Serious bleeding did not take place with the 12 cases during surgery, and surgery progressed smoothly; one of patients had heavy nosebleed after surgery, however her condition was stable when received active treatment. Follow-up 3 months to 2 years, nasal diseases of 12 patients recovered well and symptoms were relieved; cardiovascular events such as hemorrhage, thrombosis and so on did not occur. CONCLUSION Due to physiological function of the heart dysfunction in patients with coronary heart disease after PCI,they often accompany a number of trouble issues such as medical disorders, oral antiplatelet drugs, surgery affordability loss and increase surgical risk. Correct and effective perioperative treatments, strictly surgical indications are really necessary which can keep patients safe through perioperative period.
Coronary Disease ; Endoscopy ; Epistaxis ; Humans ; Nasal Septum ; surgery ; Nasal Surgical Procedures ; Nose Deformities, Acquired ; Nose Neoplasms ; surgery ; Paranasal Sinuses ; Percutaneous Coronary Intervention ; Retrospective Studies ; Treatment Outcome

Objective To investigate the effects of cardiopulmonary bypass (CPB) on the expression of tight junction protein occludin in rat lung tissues.Methods Twenty adult male Sprague-Dawley rats were randomly divided into 2 groups (n=10 each): sham operation group (group S) and CPBgroup.The rats underwent 1 h of CPB and were observed 2 h later in group CPB.The lung water content,neutrophil percentage and protein concentration in bronchoalveolar lavage fluid (BALF) were measured.The expression of occludin in lung tissues was detected by immunohistochemistry and Western-blot analysis.The ultrastructure of alveolar epithelial barrier was observed with transmission electron microscope.Results Compared with group S,the lung water content,protein concentration in BALF and neutrophil percentage were significantly increased (P ＜ 0.05),the expression of occludin in lung tissues was significantly down-regulated (P ＜ 0.05) and the damage to alveolar epithelial barrier was aggravated in group CPB.Conclusion The expression of occludin in lung tissues is down-regulated and the damage to alveolar epithelial barrier is induced after CPB,which may be one of the important factors in acute lung injury induced by CPB.

[Summary] To investigate the effect of gliclazide on the proliferation and differentiation in MC3T3-E1 cells exposed to normal glucose concentration by applying CCK-8 and RT-PCR. Gliclazide improved the proliferation and stimulated COL-I, OPN and Runx2 mRNA expression in MC3T3-E1 cells, the best concentration of gliclazide was 20μmol/ L, the expression of COL-1 and OPN mRNA had a significant positive correlation with Runx2 binding activity.

Objective To investigate the effect of penehyclidine hydrochloride on brain injury induced by cardiopulmonary bypass (CPB) in rats. Methods Thirty adult male SD rats were randomly divided into 5 groups ( n = 6 each): sham operation group (group S), CPB group, and low, median and high dose penehyclidine hydrochloride groups (group PL, PM , PH). Penehyclidine hydrochloride 0.2, 0.6 and 2.0 mg/kg were added to the priming solution in group PL, PM and PH respectively, while the equal volume of normal saline was added instead in group S. Blood samples were obtained at 2 h after termination of CPB to determine the plasma concentrations of neuron specific enolase (NSE) and S-100β protein. The brain tissues were taken to observe the ultrastructure of hippocampal neurons with electron microscope. Results The concentrations of NSE and S-100β protein were significantly higher in the other groups than in group S, while lower in group PM and PH than in group CPB and PL( P＜ 0.05). The S-100β protein concentration was significantly lower in group PH than in group PM( P ＜ 0.05). The damage to hippocampal neurons was significantly attenuated in group PM and Ps. Conclusion Penehyclidine hydrochloride 0.6 or 2.0 mg/kg can reduce the CPB-induced brain injury in a dose-dependent manner in rats.

Objective To design nursing occupational safety and health curriculum reasonably.Methods With a semi-structured interview outline,focus group interviews among eight clinical nursing and nursing education specialists were conducted to develop the occupational safety and health curriculum design.Results It was important to set up nursing occupational safety and health curriculum.The main object,contents,teaching and evaluation methods were identified.Conclusions Occupational safety and health curriculum can be involved in nursing curriculum system to reduce occupational hazards and maintain physical and mental health of nursing students.

Objective To analyze cancer incidence and mortality in Fujian in 2012 and to provide sci-entific basis for tumor prevention .Methods In accordance with the methods and criteria of data quality control made by NCCR,7 regristries data qualified from 9 submitted regritries in Fujian after data assessment were mer-ged and analyzed.Results In 2012,the cancer incidence rate was 251.44/105 (308.44/105 in male and 193.03/105 in female),age standardized incidence by Chinese standard population (ASR China)and by world standard population(ASR world)were 205.15/105 and 201.11/105.The cumulative incidence(0~74 age)was 23.51%.The mortality rate was 161.85/105(220.87/105 in male and 101.37/105 in female).ASR China and ASR world were 128.54/105 and 127.70/105 the cumulative incidence(0~74 age)was 15.04%.The age-spe-cific incidence and mortality reached maximum value in 75 ages and 80 ages respectively .The top 5 cancer inci-dences were lung cancer ,stomach cancer ,liver cancer ,esophagus cancer and breast cancer .The top 5 cancer mor-tality were lung cancer ,liver cancer ,stomach cancer ,esophagus cancer and colorectum cancer .Conclusion Di-gestive malignancies ,lung cancer ,and breast cancer in female were the most frequent in Fujian province ,and the prevention and control for those cancers should be enhanced .

Objective To evaluate the effect of hemorrhagic shock factor on the pharmacokinetics of rocuronium in pigs.Methods Sixteen pathogen-free Bama miniature pigs of both sexes,aged 3-5 months,weighing 22-25 kg,were divided into 2 groups (n=8 each) using a random number table:control group (group C) and hemorrhagic shock group (group HS).In group C,rocuronium 3.78 mg/kg was injected via the auricular vein.In group HS,the animals were subjected to volume-controlled hemorrhage,about 40％ of blood volume was withdrawn from the left femoral artery over 15 min (30 ml/kg),and rocuronium 3.78 mg/kg was injected via the auricular vein after the model was successfully established.At 0,2,4,7,10,15,20,30,60,120,180,240,300,360 and 420 min after rocuronium injection,blood samples were collected from the internal jugular vein for determination of the plasma concentration of rocuronium by high-performance liquid chromatography-tandem mass spectrometry.The pharmacokinetic parameters of rocuronium were calculated.Results Compared with group C,the plasma concentration of rocuronium was significantly increased at 20 and 60-420 min after rocuronium injection,the elimination half-life and mean residence time were prolonged,and the plasma effect-site equilibration rate constant was decreased in group HS (P＜0.05).There was no significant difference in the maximal concentration and area under the concentration-time curve between the two groups (P＞ 0.05).Conclusion The elimination of rocuronium is slower in a pig model of hemorrhagic shock.