Cloning and identification of differentially expressed genes or proteins is helpful not only for finding the functions of genes and proteins, but also for discovery of the mechanism of some diseases. Some methods have been developed for screening differentially expressed genes, such as differential display RT-PCR (DDRT PCR), subtractive hybridization (SH), DNA chip technique, and serial analysis of gene expression (SAGE). In subtractive hybridization, there have advanced three improved methods which include representational difference analysis (RDA), suppression subtractive hybridization (SSH), and full-length-gene-obtainable subtractive hybridization. For obtaining differentially expressed proteins, scientists have only two choices so far. One is two-dimentional gel electrophoresis. The other is phage display antibody repertoire library technique. Since all of the methods above have their own advantages and disadvantages, they should be used according to different needs.screening, differentially expressed genes, differentially expressed proteins

OBJECTIVE:To provide reference for the development of clinical pharmacists’career and the establishment of le-gal system in China. METHODS:The background,progress and situation of applying for“pharmacists’health care provider sta-tus”in the United States were introduced. The reasons for achieving provider status in California,Washington,and Oregon were summed up in aspects of politics,economics and education. The chance and challenge for achieving provider status at federal level were also discussed. Based on the development of clinical pharmacists in China,the suggestions were put forward for the improve-ment of legal system of clinical pharmacists in China. RESULTS & CONCLUSIONS:“Health care provider status”in the United Stated were recognised by 3 states with local developed economics,high-level education,and collective efforts of pharmacy organi-zations. Multiple national pharmacy organizations as American Pharmacists Association,American Society of Health-System Phar-macists and American Pharmacy College Society are working together toward provider status at the federal level. Our country should pay attention to related legal system construction,establish perfect and definite clinical pharmacists legal system as soon as possible to provide legal guarantee for career development of clinical pharmacists in China.

Based on the realistic circumstances at home and successful experiences from abroad,this paper holds the idea to introduce medical liability insurance into medical market to relieve the currently tense physician-patient conflict and analyzes the role and function of medical liability insurance.Meanwhile,some problems and countermeasures are also explained in the functioning process of medical liability insurance.

Objective:To measure serum levels of salusins and catestatin and analyze their correlation in patients with essential hypertension (EH) .Methods :A total of 90 EH patients were selected as hypertension group .According to blood pressure level ,they were further divided into hypertension stage 1 group (n=31) ,hypertension stage 2 group (n=30) and hypertension stage 3 group (n=29) .Another 40 normotensive subjects undergoing physical examina‐tion were selected as normal control group .Enzyme linked immunosorbent assay (ELISA) was used to measure ser‐um levels of salusins and catestatin , and the correlation between serum levels of salusins and catestatin was analyzed . Results :Compared with normal control group ,there were significant reductions in serum levels of salusins [ (3.01 ± 0.66) ng/ml vs .(1.44 ± 0.42) ng/ml ,(1.35 ± 0.89) ng/ml ,(1.41 ± 0.32) ng/ml] and catestatin [(132.24 ± 7.55) ng/ml vs .(89.22 ± 6.12) ng/ml vs .(82.51 ± 8.37) ng/ml ,(83.34 ± 4.47) ng/ml] in hypertension stage 1 ,stage 2 and stage 3 group , P<0.01 all;compared with stage 1 group ,there were significant reductions in serum catestatin levels in stage 2 and stage 3 group (P<0.01 both) ,but there were no significant difference in serum salusins level a‐mong three groups of hypertension ( P>0.05 all) .Pearson correlation analysis indicated that serum salusins level had no correlation with catestatin level ( r=0.363 , P>0.05) in normal control group ,while serum salusins level was significant positively correlated with catestatin level (r=0.723 ,P<0.01) in hypertension group .Conclusion:Serum levels of salusins and catestatin significantly reduce and they is positive correlation in patients with hyperten‐sion .Along with blood pressure level rises ,serum catestatin level reduces .

Objective: To investigate changes of plasma glucagon level in patients with heart failure (HF) caused by coronary heart disease (CHD) and its clinical significance. Methods: A total of 30 HF patients caused by CHD were selected as HF group, another 30 healthy subjects with corresponding age and gender were regard as normal control group. HF patients received comprehensive therapy of enhancing myocardial contractility, diuretic and vasodilator of 7~10d according to Chinese diagnostic and treatment guideline of chronic heart failure. Changes of glucagon level before and after treatment were observed. Results: Before treatment, plasma level of glucagon in HF group was significantly higher than that of normal control group [(205.67±120.22) ng/L vs. (90.53±20.5) ng/L, P<0.05]. After treatment, plasma level of glucagon [(120.42±30.33) ng/L] significantly decreased than before treatment (P<0.05) in HF group. Conclusion: Plasma glucagon significantly increases in patients with heart failure and gradually decreases to near normal level after treatment. Glucagon level may be regard as one of indexes judging patients’ condition.

Objective To study on effects of Alprostadil on 24-hour urinary albumin(U-Alb)in patients with diabetic nephropathy.Methods:38 patients with diabetic nephropathy were included in the study.The test group involving 20 patients were grouped into group A(pristine diabetic nephropathy) and group B(clinical diabetic nephropathy)randomly according to urinary albumin.The control group including 18 patients were grouped into two groups according to the same way.The subjects in both groups have the same therapy in terms of control of blood glucose,blood pressure,plasma lipids,and other therapy methods.Besides,the test group was injected Alprostadil(凯时)10ug from vein,one time a day,14 day continuously,but Alprostadilas well.Results After the treatment of two groups,the level of the urinary albumin decreased obviously compared with the index before treathent,and decreased significantly compared with normal control group.Conclusion Alprostadil can lower urinary albumin with diabetic nephropathy.

AIM To evaluate the beneficial effects of isoliensinine on paraquat(PQ)-induced acute lung injury and pulmonary fibrosis and explore the mechanism of its action. METHODS PQ (45 mg·kg-1, ip)-induced acute lung injury and PQ (100 mg·kg-1, ig)-induced pulmonary fibrosis were prepared. At 8, 24 and 48 h after PQ administration, the effects of isoliensinine (20 mg·kg-1, ig, 3 times a day, from 24 h before PQ administration to the end of experiment) on activity of superoxide dismutase (SOD), alkaline phosphatase (ALP) and the content of malondialdehyde (MDA) in plasma and bronchoalveolar lavage fluid (BALF) of acute lung injury groups were evaluated respectively. On the 14 d following PQ ingestion, the effects of isoliensinine (10, 20 and 40 mg·kg-1, ig, twice a day, from 24 h before PQ administration to the end of experiment) on hydroxyproline content, transforming growth factor β1 (TGF-β1) and matrix metalloproteinase-2 (MMP-2) expressions and the histopathological changes in lung tissues of pulmonary fibrosis groups were observed. RESULTS In the acute lung injury model, isoliensinine (20 mg·kg-1) significantly increased SOD activity, and decreased MDA content and ALP activity, as well as ameliorated the histopathological damage of lung tissue compared with PQ group. However, the indexes mentioned above in isoliensinine alone group did not change obviously compared with normal saline group. In the pulmonary fibrosis model, isoliensinine (10, 20 and 40 mg·kg-1) resulted in a dose-dependent decrease of hydroxyproline content compared with PQ group [(2.11±0.21), (1.94±0.24) and (1.89±0.26), respectively, vs (2.44±0.33) mg·g-1 wet tissue]. The expressions of TGF-β1 and MMP-2 in the lung tissue of the isoliensinine 40 mg·kg-1+PQ group were significantly less than those of the PQ group. Furthermore, isoliensinine could improve the histopathological changes of fibrosis as comparison with PQ group. CONCLUSION Isoliensinine has protective effects on PQ-induced acute lung injury and pulmonary fibrosis.

Objective To observe the effect of Jian Pi Hua Shi Ke Li(JPHSKL) on IBS‐D ,gastrointestinal peptide hormones of rats 5‐HT ,and explore the therapeutic mechanism .Methods 60 clean‐level Wistar rats were used in the experiment .The rats were randomly divided into normal group ,model group ,JPHS low ,middle ,high and Deshute group(10 rats in each group) .Used gavage senna and restraint stress to establish D‐IBS rat model .The drug contrations of JPHSKL low ,midle ,high group were 1 .5 ,2 .5 ,4 .0 g/mL ,3 mg/mol in Deshute group .Once each day ,for 2 weeks ,the other groups were nomol group without any treatment and mod‐el group which building successful .Immunohistochemical SABC method to observe 5‐HT level of colonic specimens .Results The 5‐HT level in colonic tissue of the rats in IBS‐D model were obviously higher than the nomal group(P<0 .01) ,and the 5‐HT level in colonic tissue in JPHSKL treated group were obviously lower than the mold group (P<0 .05) .Among them ,the JPHSKL high group had significant effect and there was no obviously difference with Deshute group (P>0 .05) .Conclusion JPHSKL can im‐prove gastrointestinal function of IBS‐D model rats ,effect mechanism is relating to decrease the level of 5‐HT in colonic mucosa .

Objective To study the sensitivity and reliability of rat skin anaphylactoid test method, detect and evaluate the anaphylactoid reaction of traditional Chinese medecine injection. Methods The condition of rat skin anaphylactoid test was optimized by studying the influencing factors of sensitivity and reliability of test with C48/80 as a tool drug, Tween80, endotoxin, China cobra toxin, trichosanthin injection, Tanreqing injection and Xuesaitong injection were investigated. Results The best conditions of rat skin anaphylactoid test was as follows:intrademal inject the drug with insulin syringe, 50-100μL per point, immediately inject 0.5%Evans blue dye 1 mL though caudal vein, 15 min later, kill the rat by carotid artery bleeding, clip dorsal skin to do the test. With this method, Tween80, endotoxin, China cobra toxin and trichosanthin injection all can induce blue stain in rat skin. Tanreqing injection showed no blue stain at the clinical dose. Xuesaitong injection although can induce blue stain in rat skin at the clinical dose, but the results cannot exclude the interference of its pharmacological function. Conclusion The method is simple with short test cycle, less dose of test drug, high detection sensitivity and good reproducibility, but some drug can show false positive result due to its own property.

Objective:To observe change of peripheral serum levels of soluble ST 2 (sST2 ) and copeptin in patients with chronic heart disease (CHF) ,and explore their diagnostic value for heart failure . Methods :A total of 80 CHF patients ,who hospitalized in our department of cardiology from Jun 1st ,2015 to Dec 1st ,2015 ,were selected as CHF group .Meanwhile ,another 30 patients with cardiac neurosis or arrhythmia were regarded as normal cardiac function group .According to NYHA classification ,CHF group was further divided into class II group (n= 22 ) , class III group (n=32) and class IV group (n=26) .Serum sST2 and copeptin levels were measured and compared a‐mong all groups ,and their correlation was analyzed .Results:Compared with normal cardiac function group ,there were significant rise in serum levels of sST2 [(0.27 ± 0.09) ng/ml vs .(1.18 ± 0.81) ng/ml] and copeptin [(1239.64 ± 229.98) pg/ml vs .(1987.16 ± 426.84) pg/ml] in CHF group , P< 0.01 both .Compared with class II group , there were significant rise in serum levels of sST2 [ (0.46 ± 0.17) ng/ml vs .(0.77 ± 0.18) ng/ml vs .(2.31 ± 0.29) ng/ml] and copeptin [ (1625.61 ± 522.13) pg/ml vs .(2103.15 ± 340.74) pg/ml vs .(2194.31 ± 332.97) pg/ml] in class III and class IV group ,and those of class IV group were significantly higher than those of class III group , P<0.05 or <0.01 .Pearson product‐moment correlation analysis indicated that serum sST2 level was significant posi‐tively correlated with serum copeptin level in CHF group (r=0.66 ,P=0.005) .Conclusion:Serum sST2 and copep‐tin levels are closely associated with severity of heart failure .They can be used as markers for heart failure ,which is worth extending .