Objective To investigate whether the Notch-1 signaling pathway is involved in the acquisition of the epithelial-mesenchymal transition (EMT) phenotype of gefitinib-acquired resistant lung cancer cells.Methods The PC9 cell line (harboring EGFR exon 19 deletion) and PC9/AB2 cells (gefitinibacquired resistant PC9 cells) were used.siRNA targeting Notch-1 eukaryotic expression vector (siNotch-1) was constructed and PC9/AB2 cells were transfected with siNotch-1.The protein expression of EGFR,Akt,Erk,Notch receptors and ligands,TGF-β receptors,E-cadherin,Vimentin,and Snail were detected by Western blot assay.DNA sequencing and fluorescence in situ hybridization (FISH) analysis were processed to detect mutation of EGFR exon 20 and MET amplification,respectively.For cytotoxicity assay,cell viability was assessed by Cell Counting Kit 8.Results Gefitinib resistant cell line PC9/AB2 had no evidence of MET amplification or EGFR T790M mutation.The expression of Notch-1 was upregulated in gefitinib resistant PC9/AB2 cells compared with that in gefitinib-sensitive PC9 cells.There were no significant protein expression differences of other Notch receptors,Notch ligands or TGF-β receptors between both paired cell lines.Western blot results showed that protein expression of E-cadherin was greatly reduced in PC9/AB2 cells,while elevated levels of Vimentin and Snail were observed.A significant reduction of the expression of Snail and Vimentin in Notch-1 siRNA transfected PC9/AB2 cells with increased E-cadherin expression was found by Western blot assay.PC9/AB2 cells displayed a round like cell morphology after Notch-1 siRNA transfection.Silence of Notch-1 decreased colony-formation ability but enhanced sensitivity of gefitinib on PC9/AB2 cells.Conclusion Notch-1 might play a novel role in acquired resistance to gefitinib,which could be reversed by inhibiting Notch-l.

[Objective] To observe the effect of Allergic Rhinitis Oral Liquid (AROL) on perennial allergic rhinitis (PAR) and to explore its therapeutic mechanism. AROL, composed of modified Guizhi Tang and Yupingfeng San, has the actions of strengthening spleen and lung, replenishing Qi and consolidating superficies and dispelling pathogens and dredging orifices. [Methods] One hundred and twenty-nine cases of confirmed PAR were randomized into Group A (n=43), Group B (n=43) and Group C (n=43). Group A was treated with AROL, Group B with Biyankang Tablets and Group C with normal saline. Three groups were treated for 4 weeks as one treatment course, and another treatment course was carried out one week after interval. Therapeutic effect were observed after treatment and serum levels of interleukin-4 (IL-4) and specific IgE antibody were detected. [Results] The total effective rate was 93.02% in Group A (P

Objective To investigate the changes in expression of hippocampal neuronal gap function protein connexin 36 (Cx36) induced by postoperative cognitive dysfunction (POCD) in aged rats.Methods Ninety male SD rats aged 20 months weighing 500-600 g were randomly divided into 3 groups ( n =30 each):group normal control (group C); group sham operation (group S) and group POCD.POCD was induced by splenectomy.Cognitive function was assessed by using open field test and maze test on the 1st,3rd and 7th day after operation (T1,T2,T3 ).The ultrastructure of gap junction in the CA1 region of hippocampas was examined with thin-section transmission electron microscope.The expression of Cx36 was detected by immuno-histochemical method.Results POCD significantly decreased the number of grid cross,the rearing and correct responses and increased the time the animab spent in the central square and total reaction time at T1 in group POCD as compared with group C.Cx36 expression was significantly decreased at T1 in group POCD as compared with group C.The ultrastructure of gap junctions underwent significant change at T1 in group POCD.Conclusion Hippocampal neuronal Cx36 may be involved in the cognitive dysfunction after splenectomy in aged rats.

CBS(Case Based Study)is a new teaching method,which is based on the case given.It requires students to answer questions according to the case. In the teaching,the students who are divided into groups,seek for resolvent by themselves. They should combine the information provided in the case,find the key answer and the relationship.The students gain the knowledge by reviewing and studying in the textbook,then analyse and solve questions with them. The application of CBS could make knowledge more relatively and systematic, moreover,it also make study more actively and initiatively.

Objective To evaluate the effects of dexmedetomidine or dezocine alone or the combination of the two agents on the emergence agitation in patients undergoing thoracic surgery.Methods Eighty ASA Ⅰ or Ⅱ patients,aged 18-64 yr,weighing 48-75 kg,scheduled for elective thoracic surgery under general anesthesia,were randomly divided into 4 groups (n =20 each):control group (group C),dexmedetomidine group (group DEX),dezocine group (group DEZ) and dexmedetomidine + dezocine group (group DEX + DEZ).In group DEX,dexmedetomidine 0.5 μg/kg was injected intravenously at 15 min before induction of anesthesia,followed by continuous infusion of dexmedetomidine at 0.4 μg· kg-1 · h-1 until beginning of chest closure.Dezocine 0.1 mg/kg was injected intravenously after beginning of chest closure in group DEZ.In group DEX + DEZ,dexmedetomidine 0.5 μg/kg was injected intravenously at 15 min before induction of anesthesia,followed by continuous infusion of dexmedetomidine at 0.4 μg·kg-1 ·h-1 until beginning of chest closure and then dezocine 0.1 mg/kg was injected intravenously.While the equal volume of normal saline was given instead in group C.Flurbiprofen axetil 50 mg was injected intravenously at beginning of skin closure in each group.Venous blood samples were collected for determination of plasma C-reactive protein (CRP),TNF-α and IL-10 levels at 10 min before induction of anesthesia (T1),before skin closure (T2),immediately after extubation (T3) and 15 min after extubation (T4).Side effects such as agitation during emergence from anesthesia were recorded.Sedation was assessed using Ramsay score.Results Compared with group C,the levels of plasma CRP and TNF-α at T2-4 and ratio of TNF-α/IL-10 were significantly decreased,the levels of IL-10 were increased at T2-4,the degree and incidence of agitation were decreased,and Ramsay score was increased in the other three groups (P ＜ 0.05).Compared with groups DEX and DEZ,the levels of plasma CRP and TNF-α at T2_4 and ratio of TNF-α/IL-10 were significantly decreased,the levels of IL-10 were increased at T2-4,and the degree and incidence of agitation were decreased in group DEX + DEZ (P ＜0.05).No side effects such as hypotension,bradycardia,respiratory depression,nausea and vomiting were observed in the four groups.Conclusion Dexmedetomidine or dezocine alone or combination of the two agents can decrease the degree and occurrence of emergence agitation and inhibit the inflammatory response simultaneously,and the combination of the two agents provides better efficacy than either alone in patients undergoing thoracic surgery.

Objective To evaluate the effect of dexmedetomidine on NF-κB activity in ventilator-induced lung injury in dogs.Methods Thirty healthy 5-year-old Beagles,weighing 10-12 kg,were randomly assigned into 5 groups (n =6 each) using a random number table:control group (group C),mechanical ventilation (MV) group and 3 different concentrations of dexmedetomidine groups (groups DEX1-3).The animals were anesthetized with pentobarbital sodium,ketamine and atropine.The rats were tracheostomized and spontaneous breathing was maintained in group C,while the rats were tracheostomized and mechanically ventilated for 4 h in MV and DEX1-3 groups.Tracheal intubation was performed in all the groups.The concentration of O2 inhaled was set at 50％,respiratory rate at 15 bpm,tidal volume at 20 ml/kg,and positive end-expiratory pressure at 2 cm H2 O.A loading dose of dexmedetomidine 0.5,1.0 and 2.0 μg/kg was infused over 20 min before intubation,followed by continuous infusion (lasting for4 h) at a rate of 0.5,1.0 and 2.0μg·kg-1 ·h-1 in DEX1-3 groups,respectively.Blood samples were taken from the femoral artery at baseline state,and 1,2 and 4 h of MV for detection of PaO2.The animals were sacrificed after 4 h of MV.The lungs were removed for determination of W/D lung weight ratio (W/ D ratio),myeloperoxidase (MPO) activity,NF-κB p65 expression (by Western blot) and TNF-α mRNA expression (by RT-PCR) in lung tissues.Results Compared with group C,W/D ratio,MPO activity,and expression of NFκB p65 and TNF-α mRNA were significantly increased in group MV (P ＜ 0.05).Compared with group MV,MPO activity,and expression of NF-κB p65 and TNF-α mRNA were significantly decreased,and no significant change was found in W/D ratio in DEX2 and DEX3 groups (P ＜ 0.05).Conclusion Dexmedetomidine can inhibit NF-κB activity and reduce inflammatory responses,thus improving ventilator-induced lung injury in dogs.

To investigate formation mechanism of secologanic acid sulfonates in sulfur-fumigated buds of Lonicera japonica, secologanic acid was enriched and purified from the sun-dried buds of L. japonica by various column chromatography on macroporus resin HPD-100, silica gel and ODS. The stimulation experiments of sulfur-fumigation process were carried out using secologanic acid reacted with SO2 in the aqueous solution. The reaction mechanism could be involved in the esterification or addition reaction. The present investigation provides substantial evidences for interpreting formation pathway of secologanic acid sulfonates in sulfur-fumigated buds of L. japonica.
Alkanesulfonates ; chemistry ; Carboxylic Acids ; chemistry ; Chromatography, High Pressure Liquid ; Flowers ; chemistry ; drug effects ; Lonicera ; chemistry ; drug effects ; Models, Chemical ; Molecular Structure ; Sulfur ; chemistry ; pharmacology ; Sulfur Dioxide ; chemistry ; Water ; chemistry

Tibetan medicine Asteris Flos is the flowers of Aster souliei, A. flaccidus, and A. asteroides, with the function of clearing away heat and toxic matter, relieving cough, and removing phlegm. In order to control the quality of Asteris Flos, the morphological and chemical methods were established for identification of three origins. The morphological features of three species were described and photographed, and the microscopic characteristics of three drug powders were also described in detail and pictured. The results showed that three origins of Asteris Flos could be easily distinguished by their macro- and micro-morphologic features, and a key for distinguishing the three origins was given. Moreover, a TLC method, with apigenin-7-O-β-D-glucurono pyranoside and chlorogenic acid as chemical references, was also established for the identification of three origins. The results showed that the TLC chromatograms of the flowers of A. souliei and A. flaccidus were very similar, but different from that of A. asteroides. The established macroscopic, powder microscopic and TLC methods for identification of three origins of AF were simple, accurate, and reproducible, and also effective and easy to operate.
Aster Plant ; anatomy & histology ; chemistry ; cytology ; Chlorogenic Acid ; analysis ; Chromatography, Thin Layer ; methods ; Flowers ; anatomy & histology ; chemistry ; cytology ; Medicine, Tibetan Traditional

Adult ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Glomerulonephritis ; complications ; Humans ; Kidney Failure, Chronic ; drug therapy ; Male ; Middle Aged ; Phytotherapy ; Powders

Objective To develop an effective way to evaluate the accurate platelet count in a patient with anticoagulants-induced pseudothrombocytopenia (PTCP).Methods It was studied that various anticoagulants effect on the platelets count for an infrequent patient with anticoagulants-dependent PTCP. When vitamin B6,aminophylline,gentamicin and amikacin were separately added to four anticoagulated blood samples from anticoagulants-dependent patient within 15 min after blood withdrawal,platelets count and morphological changes of blood cells after 4 hours of incubation at room temperature were investigated. The best anti-aggregating agent and its optimal concentration among them were explored.Results The four anticoagulants all could not inhibit the aggregation of the patient's platelets.Only amikaein among the above anti-aggregating agents can prevent and dissociate the aggregation of platelets without apparent morphological changes of blood cells and the platelet counts was stable within 4 hours after blood drawn when amikacin was added either before or after blood sampling.With increasing the concentration of amikaein,the platelet counts increase and then tend to be stable.The optimal concentration of amikacin is 5 mg/ml blood.Conclusions The supplementation of amikaein either before or after blood sampling is a useful method for the diagnosis anticoagulants-dependent PTCP and for the eva/uation of platelet counts in infrequent patients with anticoagulants-dependent PTCP.