This study was aimed to investigate the clinical effect of tetramethylpyrazine in combination with ipratropium bromide in the treatment of bronchial asthma and its effects on inflammatory factors and leptin. According to inclusion criteria, a total of 124 bronchial asthma cases were included and randomly divided into two groups, which were the control group (n = 62) and the observation group (n = 62) with the random number table. Besides conventional treatments, the control group was given ipratropium bromide; while the observation group was given tetramethylpyrazine in combination with ipratropium bromide. Then, comparisons were made between two groups on the therapeutic efficacy rate, pulmonary function, inflammatory factor, leptin, and incidence of adverse reactions. The results showed that the total therapeutic efficacy rate was 96.8% in the observation group, which was obviously higher than that of 87.1% in the control group (P < 0.05). In comparison with the control group, the posttreatment pulmonary function indexes PEF and FEV1/FVC increased, inflammatory factors IL-6 and IL-4 decreased, and IL-12 increased in the observation group with statistical significance (P < 0.05). One week, 2 weeks and 4 weeks after the treatment, the level of leptin in the observation group was significantly lower than that in the control group (P < 0.05). During the treatment, there were no severe adverse reactions. The incidence of adverse reactions in the observation group was 11.3%, which was statistically the same with that of 14.5% in the control group (P > 0.05). It was concluded that tetramethylpyrazine in combination with ipratropium bromide was effective for bronchial asthma treatment, which can significantly increase curative efficacy, improve pulmonary function, alleviate inflammatory reaction and reduce leptin level with minor adverse reactions. It required further studies in the clinical practice.

According to the constructivism approach, instructors have to adapt to the role of fa-cilitators but not teachers. Whereas a teacher gives a didactic lecture that covers the subject matter , a fa-cilitator helps the learner to get to his or her own understanding of the content. In the former scenario the learner plays a passive role and in the latter scenario the learner plays an active role in the learning pro-cess. Under the guidance of this theory, a multi-dimension teaching model based on classroom teaching, network platform and innovate experiments has been established in the course of basis of clinical labora-tory. It has been found that this model is conducive to raising students' interests in learning and to culti-vating student's comprehensive quality.

Objective To investigate the molecular mechanisms of PLCεin regulating the invasion and migration of human bladder cancer cells in vitro.Methods After cells treated with recombinant adenovirus , the migratory/in-vasive abilities of T24 cells were explored by wound healing and Transwell chamber cell migration and invasion as -say;RT-PCR was used to detect the mRNA levels of PLCε;The protein levels of PLCε,PKCα,PKCβ, TBX3 and E-cadherin were determined by Western blot;QRT-PCR was used to detect the mRNA levels of TBX3 and E-cad-herin.Results It was confirmed by digesting and sequencing that the recombinant adenovirus had been constructed successfully .The expression of PLCε mRNA and PLCε protein were both decreased after the infection of Ad-shPLCε.Wound healing and Transwell chamber cell migration/invasion assay showed that Ad-shPLCε treatment could inhibit the migratory and invasive activity of bladder cancer cells(P<0.05).The results of Western blot indicated that the expression of PKCα/βin membrane decreased ( P<0.05 ) , and phosphorylation level of PKCαand PKCβwas reduced .QRT-PCR and Western blot analysis demonstrated that the expression level of TBX 3 de-creased , but the expression level of E-cadherin increased .Conclusions PLCε shRNA can inhibit migratory and invasive ability of bladder cancer cells through PKCα/β/TBX3/E-cadherin pathway .

Objective To study the proliferation inhibition effect by silencing PLCε gene expression with RNA interference in BIU-87 cells. Methods The specific short hairpin RNA recombinant plasmids were constructed by gene clone technology.The expression level of PLCε protein and mRNA were detected by Western blot and RT-PCR respectively after transfected recombinant plasmids into BIU-87 cells.The influence on proliferation was check by MTT.The changes of proliferating cell nuclear antigen(PCNA)were analyzed by immunocytochemical method,and the distribution of cell cycle was analyzed using flow cytometry. Results After transfected with the specific recombinant plasmids,PCNA expression was decreased 33.08%,and the analysis of cell cycle indicated that cells of G0/G1 phase were increased comparision with(40.75±2.30)%and(40.00±1.76)0A,and its G2/M phase cells(8.16±0.51)%were decreased strikingly compared with group control(31.20±1.76)%and group NP(35.94±1.58)%.Cells were blocked at G0/G1 phase,the cell proliferation was inhibited obviously. Conclusion PLCε may play an important role in proliferation of bladder cancer cells,which could be a potential target of biological treatment on bladder cancer in the future.

OBJECTIVE To establish characteristic chromatogram of Yao medicine Kadsura longipedunculata and the method for the content determination of its main component anwulignan， and evaluate the anti-inflammatory activity of anwulignan. METHODS HPLC method was performed with acetonitrile-0.5% phosphoric acid solution as the mobile phase for gradient elution. The characteristic chromatogram of K. longipedunculata was established and similarity was evaluated by Similarity Evaluation System for Chromatographic Fingerprint of TCM （2012 edition）. The content of anwulignan in K. longipedunculata was determined. Lipopolysaccharide induced RAW264.7 macrophages were selected as inflammatory cell model to investigate the effects of anwulignan on the levels of tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β） and IL-6. RESULTS The similarities of characteristic chromatogram for 10 batches of K. longipedunculata ranged 0.901-0.994， and 9 common peaks were determined； 3 components were identified， such as changnan schisantherin E， kadsulactone A， anwulignan. The contents of anwulignan were （0.72±0.05）-（1.21±0.03） mg/g（n＝3）. Anwulignan of 0.125-0.5 μg/mL greatly decreased the levels of TNF-α， IL-1β and IL-6 in the supernatant of inflammatory model cells （P＜0.05 or P＜0.01）. CONCLUSIONS HPLC characteristic chromatogram of K. longipedunculata and the method for the content determination of anwulignan are all established， and anwulignan may be the active ingredient of anti-inflammatory effect in K. longipedunculata.