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

36 rabbits were divided into control and experimental groups.In the experimen-tal group,the left renal artery was ligated or the left renal artery and vein wereligated simultaneously.All the animals were alive.After 3—8 months of the opera-tion,the rabbits were killed.The establishment of collateral circulation and theanatomical,histological and histochemical changes of the kidny were observed.The resultsobtained were as follows:1.Ligation of the left renal vessels causes atrophy of theleft kidney on account of the failure of collateral circulation establishment.Theright kidney of the experimental group showed obviously compensatory hypertrophy.The weight of the compensatory hypertrophied kidneys obviously increased.2.Thecompensatory hypertrophic growth of kidneys is due to the hypertrophy of existingn(?)phrons and the hyperplasia connective tissue.3.comparison of the controls,inthe JGI,PAS and AlP reactions with the compensatory hypertrophied kidneys,there were no striking differences.The results suggest that the compensatory hyper-trophied kidneys were normal at least on the renin secretion and reabsorption func-tion.

After MC3T3-E1 cells were treated with 1,10,and 20 μmol/L glibenclamide for 48 h,the proliferation rate of cells was detected by CCK-8 assay.Flow cytometry was used to test cell apoptosis.The mRNA expressions of collagen I (COL-1) and osteopontin (OPN) were tested by realtime fluorescence quantitative PCR.Western blot was used to detect the expression levels of apoptosis-related proteins Bax and Bcl-2.The results showed that compared with the control group,the proliferation rate of MC3T3-E1 cells was gradually increased (P＜0.05),the apoptosis rate decreased (P ＜ 0.05),the expressions of COL-1 mRNA,OPN mRNA,and Bcl-2 protein were progressively raised (P＜0.05),and the expression of Bax protein were gradually decreased (P＜0.05) along with increasing concentration of glyburide.It suggested that glibenclamide could promote the proliferation and differentiation of MC3T3-E1 cells in high glucose and may inhibit apoptosis in a concentration-dependent manner within a certain range.

OBJECTIVE:To investigate the mechanism of cross allergy reaction during the application of β-lactam antibiotics, and to provide reference for rational drug use in clinic. METHODS:Based on study experience of author in UIC and its affiliated hospital during advanced study,according to the experience of drug use safety management in patients allergic to β-lactam antibiot-ics from Rush University Medical Center,the mechanism of cross allergy reaction during the application of β-lactam antibiotics was summarized,and the disposal procedure for patients allergic to β-lactam antibiotics in the Affiliated Hospital of UIC was intro-duced. RESULTS:The principal reason for cross allergy reaction induced by β-lactam antibiotics were same or similar side chains between drugs. Cross allergy reaction occurred when IgE recognized these side chains. The disposal procedure for patients allergic to β-lactam antibiotics in the Affiliated Hospital of UIC included that the indication of β-lactam use was evaluated;standard penicil-lin skin testing according to evaluation results,anti-infection treatment by Grade challenge β-lactam antibiotics and course and rap-id drug tolerance induction. CONCLUSIONS:The disposal method for patients allergic to β-lactam antibiotics in the Affiliated Hos-pital of UIC can provide new thought for domestic clinical pharmacists in rational drug use among the patiens with reported aller-gies to special group as pregnant women,children.

Objective To investigate the effects of penehyclidine hydrochloride on blood-brain barrier in a rat model of cardiopulmonary bypass ( CPB) . Methods Sixty adult male SD rats, aged 4-6 months, weighing 320- 370 g, were randomly divided into 5 groups ( n = 12 each) : sham operation group (group S), CPB group, and low-, median- and high-dose penehyclidine hydrochloride groups (groups LP, MP and HP). The animals were anesthetized with intraperitoneal 10% chloral hydrate 350 mg/kg, intubated and mechanically ventilated. The femoral and jugular arteries and jugular vein were cannulated. CPB was performed for 60 min. Penehyclidine hydrochloride 0.2, 0.6 and 2.0 mg/kg were added to the priming solution in groups LP, MP and HP respectively, while the equal volume of normal saline was added in group CPB. Evans blue was injected via femoral vein at 1 h before the animals were sacrificed. Six rats in each group were sacrificed, their brains immediately removed and the hippocampi isolated for determination of Evans blue content. The other rats were sacrificed and the hippocampi isolated to determine the water content and observe the ultrastructure of blood-brain barrier. Results Compared with group S, the Evans blue content and water content were significantly increased in the other groups ( P ＜ 0.05) . Compared with groups CPB and LP, the Evans blue content and water content were significantly decreased in groups MP and HP ( P ＜ 0.05) . The Evans blue content was significantly lower in group HP than in group MP ( P ＜ 0.05). The CPB-induced changes were significantly attenuated in groups MP and HP compared with groups CPB and LP. Conclusion Penehyclidine hydrochloride can protect blood-brain barrier against the CPB-induced injury and the effect is related to the dose.

ObjectiveTo investigate the effect of penehyclidine hydrochloride on acute lung injury induced by cardiopulmonary bypass (CPB) in rats.MethodsForty adult male SD rats aged 4-6 months weighing 330-420 g were randomly divided into4 groups ( n =10 each): sham operation group (group S),acute lung injury group (group ALI) and low and high dose of penehyclidine hydrochloride groups (groups PL and PH ).Penehyclidine hydrochloride 0.6 and 2.0 mg/kg were added to the priming solution in groups PL and PH,while the equal volume of normal saline was added in group ALI instead.The rats of groups ALI,PL and PH were underwent 1 h of CPB.Arterial blood samples were collected before CPB and at 2 h after CPB for blood gas analysis.The superior vera cava blood samples and lung tissues were collected at 2 h after CPB for determination of concentrations of TNF-α and IL-6,lung tissue contents of water and malondialdehyde (MDA) and activity of glutathione peroxidase (GSH-px).The pathological change of lung tissue was also examined.ResultsCompared with group S,PaO2 was significantly decreased at 2 h after CPB,plasma concentrations of TNF-α and IL-6 and contents of water and MDA in lung tissues were increased,while activity of GSH-px in lung tissues was decreased in groups ALI,PL and PH ( R ＜ 0.05).Compared with group ALI,PaO2 was significantly increased at 2 h after CPB,plasma concentrations of TNF-α and IL-6 and contents of water and MDA in lung tissues were decreased,activity of GSH-px in lung tissues was increased (P ＜ 0.05),and the pathological change was reduced in groups PL and PH.Compared with group PL,PaO2 was significantly increased at 2 h after CPB,plasma concentrations of TNF-α and IL-6 and contents of water and MDA in lung tissues were decreased,activity of GSH-px in lung tissues was increased ( P ＜0.05),and the pathological change was reduced more obviously in group PH.ConclusionPenehyclidine hydrochloride 0.6 or 2.0 mg/kg can reduce the CPB-induced lung injury in a dose-dependent manner by antioxidant and anti-inflammatory mechanism in rats.

Objective To investigate the efficiency of transduction of recombinant adenovirus-mediated human endothelial nitric oxide synthase (eNOS) into lung tissue by repeated intratracheal transfection in rats.Methods Sixty 3-4 month old male Wistar rats weighing 220-280 g were randomly divided into 2 groups:control group (group C,n =10) and eNOS gene transduction group (group T,n =50).The animals were anesthetized with intraperitoneal 10％ chloral hydrate 35 mg/kg,tracheally intubated and mechanically ventilated (VT 2.5 ml,RR 60 bpm,FiO2 1.0).Recombinant adenovirus carrying human eNOS gene was given as gift by Professor Gerard from Texas University,Southwest Medical Center.In group T 50 μl of the recombinant adenovirus in concentration of 5 × 109 PFU/ml was instilled into trachea every 5 minutes for 12 times,while in group C equal volume of vector conservation solution was instilled instead.Pulmonary arterial blood samples were obtained at 2,5,7,14 and 21 d after intratracheal transfection (n =10 at each time point) for determination of serum NO concentration.The animals were immediately sacrificed after blood sample collection for determination of expression of eNOS protein in the lung tissue and RNA.The eNOS expression in the trachea,bronchus,lung,liver,spleen and kidney was detected by immuno-histochemistry.Results The serum NO concentrations were significantly higher at all time points in group T than in group C.The eNOS expression was detected in the epithelial cells of trachea and bronchi,and endothelial cells of alveoli and pulmonary blood vessels in group T but not in group C.eNOS expression was not detected in liver,spleen and kidney at 7 d after intratracheal transfection in group T.Conclusion Human eNOS gene mediated by recombinant adenovirus was transducted into rat lung tissue with normal enzyme activity by repeated intratracheal administration without being detected in distant organs.