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 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 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 evaluate the bioavallability and bioequivalence of rabeprazole sodium enteric-coated pellets capsules. Methods:A randomized crossover design was performed in 32 healthy male volunteers. A single oral dose of 20 mg rabeprazole sodium enteric-coated pellets capsules ( test preparation) or enteric-coated shell capsules ( the reference capsules) was administrated under fed conditions. The wash period was 7 days. The blood samples were collected at different time points. The concentration of rabeprazole in plasma was determined by an LC-MS/MS method. The pharmacokinetic parameters were calculated by DAS 3. 0 software and the bio-equivalence was evaluated. Results:The maln pharmacokinetic parameters of the two formulations were shown as follows:T1/2 of (2. 20 ± 0. 83)h and(1. 951 ± 0. 515)h,Tmax of (3. 88 ± 1. 11)h and(4. 64 ± 1. 504)h,Cmax of (401. 06 ± 170. 75)ng·ml-1 and(394. 63 ± 215.64)ng·ml-1,AUC0→t of (918.42 ±427.39)ng·h·ml-1 and (994.49 ±520.73)ng·h·ml-1, and AUC0→∞ of(937.30 ± 445.13)ng·h·ml-1 and(1 011.69 ±534.77)ng·h·ml-1. The analysis showed that the maln pharmacokinetic parameters of the two formulations had no significant differences(P>0. 05) except for Tmax(P<0. 05). The relative bioavallability of rabeprazole sodium enteric-coated pellets capsules was (99. 80 ± 7. 20) %. Conclusion:Compared with the reference capsules, rabeprazole sodium enter-ic-coated pellets capsules show the property of higher dispersion degree, milder influence from food, more rapid release and absorption. The enteric-coated pellets capsules and the reference capsules are bioequivalence.

[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 dexmedetomidine on acute kidney injury in endotoxemic rats.Methods Thirty adult male Sprague-Dawley rats,aged 4-6 months,weighing 180-220 g,were randomly divided into 3 groups (n =10 each) using a random number table:control group (group C),lipopolysaccharide group (group L),and dexmedetomidine (group D).Lipopolysaccharide (LPS) 5 mg/kg was injected slowly into the femoral vein to establish the model of endotoxemic in rats anesthetized with chloral hydrate.In group D,after LPS injection,a loading dose of dexmedetomidine 7 μg/kg was injected intravenously,and 15 min later dexmedetomidine was infused for 6 h at 5 μg · kg-1 · h-1,while the equal volume of normal saline was given in L and C groups.At 6 h after the end of LPS administration,blood samples were collected from the femoral vein for determination of serum creatinine (Cr),blood urea nitrogen (BUN),tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) concentrations.At 24 h after the end of LPS administration,the animals were sacrificed and kidneys were removed for microscopic examination and for determination of the expression of tight junction proteins ZO-1 and occludin in renal tissues by Western blot.Results Compared with group C,the serum Cr,BUN,TNF-α and IL-6 concentrations were significantly increased,and the expression of ZO-1 and occluding was down-regulated in L and D groups.Compared with group L,the serum Cr,BUN,TNF-α and IL-6 concentrations were significantly decreased,the expression of ZO-1 and occluding was up-regulated,and the pathological changes of kidneys were mitigated in D group.Conclusion Dexmedetomidine can alleviate acute kidney injury in endotoxemic rats.

Objective To evaluate the effect of dexmedetomidine on acute liver injury in rats with endotoxemia.Methods Eighteen adult male Sprague-Dawley rats, aged 3-4 months, weighing 250-300 g, were randomly divided into 3 groups (n=6 each) using a random number table: control group (group C), endotoxin group (group E), and dexmedetomidine group (group D).In E and D groups, lipopo-lysaccharide 5 mg/kg was injected via the femoral vein of rats anesthetized with chloral hydrate.In group D, dexmedetomidine was infused with a 7 μg/kg loading bolus over 15 min after injection of lipopolysaccharide, followed by a 6 h continuous infusion of 5 μg · kg-1 · h-1.The equal volume of normal saline was given instead in E and C groups.After the end of administration, blood samples from the femoral vein were drawn for determination of tumor necrosis factor-alpha (TNF-α) and interleukin-1β (IL-1β) concentrations in serum (by using enzyme-linked immunosorbent assay), and alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities in serum (using the International Federation of Clinical Chemistry and Laboratory Medicine reference procedures).Liver specimens were obtained for examination of pathologic changes with electron microscope.Results Compared with group C, the serum ALT and AST activities and TNF-α and IL-lβ concentrations were significantly increased in E and D groups.Compared with group E, the serum ALT and AST activities and TNF-α and IL-1β concentrations were significantly decreased in group D.The pathologic changes of livers were obvious in group E, and were significantly reduced in group D compared with group E.Conclusion Dexmedetomidine can alleviate acute liver injury in rats with endotoxemia, and the underlying mechanism is associated with inhibition of systemic inflammatory responses.

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 evaluate the effect of penehyclidine hydrochloride (PHC) on the level of angiopoietin-1 (Ang-1) and tyrosine kinase receptor-2 (Tie-2) during endotoxin-induced acute lung injury (ALI) in rats.Methods Forty adult male Sprague-Dawley rats,weighing 180-220 g,were randomly divided into 4 groups using a random number table (n =10 each):control group (group C),ALI group,low-dose PHC group (group L-PHC) and high-dose PHC group (group H-PHC).ALI was induced with iv injection of lipopolysaccharide 5.0 mg/kg via the tail vein.In L-PHC and H-PHC groups,PHC 0.6 and 2 mg/kg were injected,respectively,via the tail vein at 1 and 24 h after lipopolysaccharide injection.The rats were sacrificed at 48 h after the initial injection of PHC to measure the lung water content,protein concentration in bronchoalveolar lavage fluid (BALF),and the expression of Ang-1,Tie-2 and phosphorylated Tie-2 in lung tissues.The morphological changes of lung tissues were observed under light microscope and the ultrastructural changes of alveolar epithelial barrier under transmission electron microscope.Results Compared with group C,the lung water content and protein concentrations in BALF were significantly increased,and the expression of Ang-1 and phosphorylated Tie-2 was down-regulated in the other three groups (P ＜ 0.05).Compared with group ALI,the lung water content and protein concentrations in BALF were significantly decreased,and the expression of Ang-1 and phosphorylated Tie-2 was up-regulated in H-PHC group (P ＜ 0.05),and no significant changes were found in the parameters mentioned above in group L-PHC (P ＞0.05).The damage to lung tissues was significantly reduced in group H-PHC as compared with group ALI.Conclusion PHC can improve the permeability of pulmonary microvascular and reduce injury to alveolar epithelial barrier,thus ameliorating endotoxin-induced ALI in rats,and the effect is dose-related and up-regulation of Ang-1 expression and inhancement of Tie-2 activity are involved in the mechanism.