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 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 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.

AIM:To explore the effect of homocysteine (Hcy)-mediated endoplasmic reticulum stress on lipid metabolism in hepatocytes.METHODS: HepG2 cells used in the study were treated with 5 mmol/L Hcy. The concentrations of Hcy, triglycerides (TGE) and cholesterol (CHO) in the cells were measured. In high methionine diet-induced hyperhomocysteinemia C57BL/6 mice, the concentrations of TGE and CHO in hepatocytes were analyzed. The mRNAs and proteins expressions of glucose-regulated protein 78 (GRP-78) and sterol regulatory element binding protein (SREBP-1) were also assessed.RESULTS: The concentrations of Hcy and lipids (TGE, CHO) in HepG2 cells at different time point were elevated after treated with 5 mmol/L Hcy (P0.05,vs 0 week). The mRNAs and proteins expressions of GRP-78 and SREBP-1 in mice at different time point after high methionine diet were higher than that at 0 week (P

OBJECTIVETo compare the cleaning effect of automatic cleaning machine and manual cleaning on coupling type surgical instruments.

METHODSA total of 32 cleaned medical instruments were randomly sampled from medical institutions in Putuo District medical institutions disinfection supply center. Hygiena System SUREII ATP was used to monitor the ATP value, and the cleaning effect was evaluated.

RESULTSThe surface ATP values of the medical instrument of manual cleaning were higher than that of the automatic cleaning machine.

CONCLUSIONCoupling type surgical instruments has better cleaning effect of automatic cleaning machine before disinfection, the application is recommended.

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 consolidate the results of special antibacterial drug rectification so as to improve the rational medi-cation of antibacterial drugs. METHODS:By using the method of retrospective analysis,the antibacterial drug application data in 2011(before rectification and initial stage of rectification),2012(middle stage thereof)and 2013(later stage thereof)were sum-marized,compared and analyzed. RESULTS:After special rectification(Jan. 2011 vs. Dec. 2013),the utilization rate of antibacte-rial drugs in perioperative period for class I incision decreased from 80.42% to 24.49%. That of antibacterial drugs in emergency treatment decreased from 53.79% to 36.23%. The examination rate of microbial samples in inpatients who would use special-grade antibacterial drugs increased from 16.12% to 94.17%. The use intensity of antibacterial drugs by inpatients decreased from 70.28 DDDs/(100 persons·d) to 27.44 DDDs/(100 persons·d). The utilization rate of antibacterial drugs by inpatients decreased from 65.99% to 46.45%. CONCLUSIONS:Special rectification has remarkable effects on rational clinical application of antibacterial drugs. Regulation and evaluation should be strengthened to maintain the effects and continuously improvement should be made.