By retrospective analysis method, the paper was summarized experience of clinical pharmacy of TCM. The purpose of the paper was given location of clinical pharmacists in Traditional Chinese Medicine(TCM), to clear their main status and functions, seek methods and ways to solve the problems in the process of work. The work was proceeding orderly. Good results have been achieved. The clinical pharmacists of TCM participate in clinical rational use of drugs to change the service mode ofindirect service for the patients in the past with the patients, doctors and nurses so as to make better service for people's life and health of Chinese medicine.

Objective To investigate the killing effect of ethacrynic acid (EA) on lung cancer A549 cells derived spheres and explore the underlying mechanism.Methods A549 spheres were cultured in serum-free medium,and the protein expression of CD133,SOX2,EpCAM and ABCG2 was detected by Western blotting.MTT assay was used to evaluate the cell viability of A549 spheres and A549 cells after treated by 1,2,5,10 and 20 mg/mL cisplatin (DDP) for 48 h.The activity of glutathione S-transferase (GST) was measured by colorimetric method after A549 spheres were treated with 10,50,100 and 200 μmol/L EA,respectively.Flow cytometry,Western blotting,real-time PCR and luciferase assay were used to analyze the levels of cellular reactive oxygen species (ROS),formation of A549 spheres,mRNA and protein expression levels of β-catenin,Sox2 and ABCG2,and promoter activity of β-catenin upon 200 μmol/L EA treated cells for 48 h.A549 sphere was infected with β-catenin adenovirus for 24 h,followed by 200 μmol/L EA treatment (in presence or absence of 5 mg/mL DDP) for 24 h.The expression of β-catenin,Sox2 and ABCG2 at mRNA and protein levels was detected by real-time PCR and Western blotting,and cell growth of A549 spheres was evaluated by MTT assay.Results The A549 spheres,with high expression of tumor stem cells markers CD133,SOX2,EpCAM and drug resistance related molecule ABCG2,and resistance to DDP at different doses,were successfully derived.After 200 μmol/L EA had treated A549 sphere for 48 h,the levels of ROS were significantly increased (P ＜ 0.05),and the mRNA and protein levels of β-catenin,Sox2 and ABCG2,and promoter activity of β-catenin were notably decreased (P ＜ 0.05).The treatment of 200 μmol/L EA enhanced the inhibitory effect on proliferation and the promoting effect on apoptosis in A549 spheres induced by 5 mg/mL DDP (P ＜ 0.05).Up-regulation of β-catenin by adenoviral infection partly reversed the effects of 200 μmol/L EA on suppressing the expression levels of β-catenin,Sox2 and ABCG2,compared to the spheres infected with blank adenovirus.Additionally,β-catenin over-expression significantly remitted the inhibitory effect of 200 μmol/L EA and 5 mg/mL DDP on the proliferation in A549 spheres.Conclusion EA exerts inhibitory effect on the proliferation and stemness of A549 spheres through suppressing GST activity and β-catenin expression,and then promotes cell apoptosis.EA might be a novel drug in treatment of lung cancer and cancer stem cells.

Objective To evaluate the importance of plasma MicroRNAs in early diagnosis of acute myocardial infarction (AMI).Methods 24 patients with AMI as the test group and 20healthy volunteers as the control group were enrolled in this study.Plasma levels of microRNA-1,microRNA-133a,microRNA-208a and microRNA-499 were detected by quantitative real time polymerase chain reaction (qRT-PCR) at 3 h,6 h,12 h,24 h,48 h,72 h after the onset of AMI.Results Plasma microRNA-1 level was greatly increased and reached the peak at 3 h after AMI,then was decreased gradually to normal level at 72 h after AMI.Plasma microRNA-133a level was significantly elevated at 6 h after AMI,reached peak at 12 h after AMI,then was decreased to normal level at 48 h after AMI.Plasma microRNA-1 and microRNA-133a levels were correlated with cTnI expression.The peak time of microRNA-1 was earlier than that of cTnI,while the peak time of microRNA-133a was the same as that of cTnI.Conclusions Increased circulating microRNA-1 and microRNA-133a may serve as potential and novel biomarkers for early diagnosis of AMI.

Objective To explore the the analytical method for the quality difference of Radix Salviae Mihiorrhizae (RSM) injection. Methods Parallel intercomparison experiment was carried out in different batches of RSM injection samples which had different degrees of adverse drug reaction. The samples were analyzed by gel size exclusion chromatog-zraphy and C18 HPLC fingerprint. Subtractive analysis was used to reveal the information of the non-low molecular weight phenolic acids. Results There were obvious differences between the results of size exclusion chromatography. The amount of the non-low molecular weight phenolic acids from different batches of the samples was also different, which was correlated with the degrees of acute toxicity in guinea pigs. Conclusion The amount of the non-low molecular weight phenolic acids varies with different batches of Radix Salviae Mihiorrhizae injection. It is suggested that non-low molecular weight phenolic acids can be used for the quality control of Radix Salviae Mihiorrhizae injection. For the first time, the perspective that the corresponding relationship between the fingerprint and their toxicities of Traditional Chinese medicine injection may be one of the most important fields is put forward.

Objective To probe into the perioperative nursing method during osterior fusion and fmation for idiopathic scoliosis. Methods The study involved 31 patients with idiopathic scoliosis treated with posterior correction nails and selective spinal fusion with hook-rod system fixation.The severe patients were treated with extensive posterior release or anterior correction and fusion posterior to release.Before the operation,the patients were administrated with mental nursing,lung function exercise,self-elongation exercise,wake-up pilot training,self-image appraisement and bed and life skills exercise.After the operation,an observation was done on the change of nervous system,propor turning the body over best clinostatism,drainage tube care,respiratory passage care,functional exercise instruction and discharge instruction. Results The operation was accomplished successfully in all the patients,which showed that the body height was increased 2-15 cm.There found no complications including loosening or breakage of the nails or sticks,nervous system injury,incisional wound infection or press~e sore. Conclusion Preoperative sufficient preparation and fine postoperative nursing are important guarantee to successful operation and fast rehabilitation.

Objective To investigate the different expressions of pathological tissue and serum microRNAs (miRNAs)in Hirschsprung disease(HSCR). Methods Pathological colon tissues and serum samples were obtained from 52 confirmed HSCR cases respectively by surgery and pathology and from 52 matched controls,respectively. An initial screening of the tissues and serum microRNA expression were performed through TaqMan Low Density Array. The candidate tissue and serum miRNAs were validated by quantitative real - time - PCR in the 20 paired array samples and extra 32 paired samples after the integration of the screening result. The bioinformatical software online including miR-base,Target Scan,PicTar and MiRanda were used to predict the target mRNA of the consistent microRNAs in the tis-sues and the serum. Results Compared with the controls,47 microRNAs were differently expressed in HSCR tissues, including 17 up - regulated miRNAs and 30 down - regulated miRNAs;32 upregulated miRNAs were also detected to be differently expressed in the HSCR serum. Among these microRNAs,miR - 218 - 1 and miR - 885 - 5p were identi-fied to have a consistent significant different expression in both tissues and the serum,which were validated as high -expressed in microarray samples and expanded 32 paired samples(miR - 218 - 1:tissue array 0. 017 58 ± 0. 002 29 vs 0. 003 37 ± 0. 000 50,P ﹤ 0. 001;tissue expanded expression 0. 013 53 ± 0. 001 74 vs 0. 004 43 ± 0. 000 60,P ﹤0. 001. miR - 885 - 5p:tissue array 0. 000 30 ± 0. 000 11 vs 0. 000 04 ± 0. 0000 08,P = 0. 027 6;tissue expanded ex-pression 0. 004 59 ± 0. 000 16 vs 0. 000 04 ± 0. 000 01,P = 0. 014 5. miR - 218 - 1:serum array 0. 769 60 ± 0. 285 50 vs 0. 045 14 ± 0. 015 07,P = 0. 015 5;serum expanded expression 1. 151 00 ± 0. 430 00 vs 0. 023 07 ± 0. 003 81,P =0. 008 7. miR -885 -5p:serum array 1. 595 00 ±0. 441 70 vs 0. 169 40 ±0. 034 46,P =0. 001 2;serum expanded expres-sion 1. 689 00 ±0. 453 00 vs 0. 146 10 ± 0. 031 24,P = 0. 001 2). Specifically,the target genes of these 2 microRNAs were RET,PLAG1 and NeuroD1,which had been reported to be directly related to HSCR. Conclusions Significantly dif-ferential expressed miRNAs exist in the pathological tissue and the serum of HSCR. MiR - 218 - 1 and miR - 885 - 5p, which showing consistent differential expression,may be involved in the pathogenesis of HSCR.

Objective To evaluate the potential of interventional regional arterial infusion for severe acute pancreatitis (SAP). Methods Regional arterial infusion with somatostatin and antibiotic was performed in 20 cases of SAP within 4 hours after diagnosis confirmed,and pressurized infusion lasted for 5 days. Another 28 cases of SAP as control group received drug treatment all over the body. The SAP clinical data such as symptom,mortality,complication rate and stay time in hospital were analysed in two groups. Results There were significant differences in remission,serum amylase,urine amylase and lipase between the regional and total infusion groups. The stay time in hospital,mortality,complication and operation rate were lower in regional infusion group than those of control group(P

Objective To explore the predictive value of ventricle late potential (LP) for arrhythmic events in the patients with Brugada syndrome. Methods Totally 43 patients with Brugada syndrome were divided into symptom group (n=24) and asymptom group (n=19).Signalaveraged electrocardiography(SAECG) was performed to analyze characteristics of LP in all subjects.The occurrence of arrhythmic events was observed in all patients during the dynamic follow-up for (33.8±9.0) months.Results There were 22 cases (91.7％) and 7 cases (36.8％) with LP positive in patients with symptom and asymptom,respectively.The incidences of arrhythmic events were 72.4％ in Brugada syndrome patients with positive LP and 14.3％ in patients with negative LP,respectively.The relative risk (RR,95％ CI) for LP prediction of the arrhythmic events was [5.1,(1.4～ 18.6)] (P =0.002). ConclusionsLP may be one of effective factors predicting arrhythmic events in the patients with Brugada syndrome.

OBJECTIVE: To determine the relationship between the number,phenotype and functional status of dendritic cells (DCs) and coronary collateral circulation (CCC) in coronary heart disease (CHD). METHODS: Forty patients with severe coronary stenosis were recruited and divided into a CCC formation group (Group A, n=22) and a non-CCC formation group (Group B, n=18). Density gradient centrifugation was applied to separate the mononuclear cells (MNCs) from coronary artery blood samples, and MNCs were cultured and proliferated in vitro. The morphology of DCs was observed under converted microscope. The number of harvested cells and DCs was counted by hematocytometer. Flow cytometry was applied to investigate the phenotype and the mean fluorescence intensity (MFI). Mixed lymphocyte reaction was used to test the function of DCs to stimulate the proliferation of T lymphocytes. Stimulation index (SI) was calculated and compared. RESULTS: (1) After in vitro proliferation, DCs were cultured successfully from the mononuclear cells from coronary artery blood samples and the morphology of DCs was not different in the 2 groups. (2) The number of mononuclear cells (MNC no) was (3.95+/-1.41)*10(6), in the CCC group and (2.76+/-0.92)*10(6) in the non-CCC group. The MNC number was significantly increased in the CCC group (P=0.003). (3) The number of DCs was (1.54+/-0.96)*10(6) in the CCC group, and (0.99+/-0.46)*10(6) in the non-CCC group (P=0.033). (4)There was no statistical significance in the percent of CD1a+, CD1a+CD80+, CD1a+CD83+, CD1a+CD86+ cells, and MFI in the 2 groups (P>0.05). (5) SI was 4.96+/-2.30 in the CCC group, whereas 2.66+/-1.04 in the non-CCC group. The SI in the CCC group increased significantly(P=0.0003). CONCLUSION In CHD patients with severe coronary stenosis, patients with CCC formation have higher number of DCs and stronger potential of T lymphocyte stimulation.
Aged ; Cells, Cultured ; Collateral Circulation ; immunology ; physiology ; Coronary Circulation ; immunology ; physiology ; Coronary Disease ; blood ; immunology ; physiopathology ; Coronary Stenosis ; blood ; immunology ; physiopathology ; Dendritic Cells ; immunology ; Female ; Humans ; Male ; Middle Aged ; T-Lymphocytes ; cytology ; immunology

Objective To explore the correlation between apolipoprotein E (ApoE) gene polymorphism and urine Alzheimer-associated neuronal thread protein (AD7c-NTP) level in patients with Alzheimer's disease (AD) and mild cognitive impairment (MCI).Methods The cognitive function of 30 AD patients (AD group),30 MCI patients (MCI group) and 30 normal controls (NC group) was evaluated by neuropsychological batteries like MMSE,the Cambridge Cognitive Examination-Chinese Version (CAMCOG-C),etc.ELISA was used to test the urine level of AD7c-NTP.The genotypes of ApoE were analyzed by the high-resolution melting assay in blood samples.Results Compared with the NC group (0.59 (0.40,0.66) ng/ml),the urine level of AD7c-NTP in the AD group (1.03(0.80,1.41) ng/ml) and the MCI group (0.69(0.53,0.91) ng/ml) was increased (Z =33.727,P ＜0.01).The urine level of AD7c-NTP in the AD group was higher than that in the MCI group (Z =8.232,P ＜ 0.05).The level of AD7c-NTP in urine was negatively correlated with MMSE and CAMCOG-C scores (rMMSE =-0.604,P ＜ 0.01;rCAMCOG-C =-0.486,P ＜ 0.01).According to receiver operating characteristic curve,the optimal cutoff point of AD7c-NTP in urine for diagnosis of patients including AD and MCI was 0.70 ng/ml,with sensitivity of 71.7％ and specificity of 83.3％,and area under the curve of 0.82 (95％ CI 0.73-0.90,P ＜0.05).There were four genotypes comprising ε2/3,ε3/3,ε3/4 and ε4/4 for ApoE gene.The frequencies of ε4 carriers were 46.7％ (14/30),23.3％ (7/30) and 23.3％ (7/30) in the AD,MCI and NC groups,respectively.There was a notable increase in urine AD7c-NTP and a significant decrease in CAMCOG-C scores in MCI patients who harbored the ApoE ε4 allele (ZAD7c-NTP =4.857,P ＜ 0.05;ZCAMCOG-C =4.284,P ＜0.05).Conclusions The urine level of AD7c-NTP was significantly increased in AD and MCI patients,the higher the level of AD7c-NTP,the more serious the cognitive impairment.The ε4 carriers exhibited higher urine level of AD7c-NTP,but worse cognitive function compared to ε4 non-carriers in the MCI group.