Objective:To study how to guide the individual dose of clopidogrel in line with the genetic testing results. Methods:Clinical pharmacists decided how to optimize the prescription of clopidogrel according to the genotype combined with the drug metabolism and drug interactions for three patients respectively with slow clopidogrel metabolism,intermediary metabolism and super fast metabolism. Results:The slow clopidogrel metabolism patient with subacute stent thrombosis after half a month of coronary stenting was switched to orally administrate with ticagrelor. The super fast metabolism patient suffered from repeatedly subcutaneous hemorrhage with antiplatelet therapy was suggested to lower the dose of clopidogrel,temporarily withdraw Maixuekang capsules and conventionally administrate with vitamin C tablets orally. The intermediary metabolism patient with late stent thrombosis co-treated with lansoprazole was suggested to increase the dose of clopidogrel or use ticagrelor instead,and when it was necessary,panxitorazole,ray Bella or the other ranitidine acid suppression drugs such as ranitidine could be considered. Conclusion:Through genetic testing and drug interactions,clinical pharmacists guide the clinical use of clopidogrel and the optimization of antiplatelet therapy.

Objective To explore the relationships between the serum and urinary levels of procollagen Ⅲ and renal injury at the early stage of hypertension in sponantously hypertensive rats. Methods 16 sponantously hypertensive rats (SHR) were divided into treated group (losartan 30mg?kg -1?d -1) and untreated group, each group containing 8 animals, and 8 Wistar-kyoto rats served as normotensive control group(WKY). The serum and urinary levels of procollagen Ⅲ and ? 2-microglobin, and urinary microalbumin level were measured by radioimmunoassay 16 weeks after treatment. Results Blood pressure, serum ? 2-microglobin level, and the urinary levels of procollgen Ⅲ, ? 2-microglobin and microalbumin in SHR were significantly higher than those in wistar-kyoto rats. Losartan could decease the levels of the indices above in SHR. Serum procollagen Ⅲ level had not significant difference among the three groups of rats, and was not correlated with urinary procollagen Ⅲ level. There were positive correlation between serum and urinary ? 2-microglobin levels, and urinary procollgen Ⅲ level was positively retated to urinary ? 2-microglobin, urinary microalbumin and systalic pressure. Conclusion The levels of urinary procollgen Ⅲ,? 2-microglobin and microalbumin could reflect the lesion of glomerulus and tubulointerstitum at the early stage of essential hypertension in SHR.

Acetylcholine ; metabolism ; Acetylcholinesterase ; metabolism ; Animals ; Brain ; enzymology ; metabolism ; Female ; Hypoxia, Brain ; physiopathology ; Ischemic Preconditioning ; methods ; Male ; Memory ; physiology ; Mice ; Random Allocation ; Reperfusion Injury ; prevention & control

To study the pharmacokinetics characteristic of loganin, ferulic acid and stilbene glucoside in rat plasma after oral administration of Bushen Tongluo formula. The plasma samples were treated by using liquid-liquid extraction technique, the concentrations were determined by HPLC-UV. Johnson spherigel C18 column (4.6 mm x 250 mm, 5 μm) was adopted and eluted with the of mobile phase of methanol-water containing 0.01% glacial acetic acid in a gradient mode, with the flow rate at 1.0 mL x min(-1), column temperature at 30 degrees C and injection volume of 10 μL. According to the findings, loganin was determined at 235 nm, ferulic acid and stilbene glucoside were determined at 320 nm, with the sample size of 10 μL. The pharmacokinetic parameters of loganin, ferulic acid and stilbene glucoside were calculated by DAS 2. 0 software as follows: C(max) was (0.369 ± 0.042), (0.387 ± 0.071), (0.233 ± 0.044) mg x L(-1); t(max) was (0.226 ± 0.022), (0.282 ± 0.031), (0.233 ± 0.044) h; t(½β) was (6.89 ± 0.20), (10.73 ± 0.11), (6.93 ± 0.09) h; AUC(0-∞) was (1.91 ± 0.36), (3.22 ± 0.52), (1.52 ± 0.33) mg x h x L(-1); AUCO(0-t) was (1.62 ± 0.33), (2.58 ± 0.43), (1.30 ± 0.30) mg x h x L(-1); CL was (20.2 ± 4.0), (1.39 ± 0.23), (31.7 ± 6.9) L x h(-1) x kg(-1), respectively. The results showed that after the oral administration with Bushen Tongluo formula, loganin, ferulic acid and stilbene glucoside showed concentration-time curves in conformity with the two compartment model, with a rapid absorption, loganin and stilbene glucoside was excreted at a moderate speed, and ferulic acid was excreted slowly (but with the highest bioavailability). Bushen Tongluo formula can main maintain plasma concentration with three administrations everyday and so is suitable to be made into common oral preparation.
Administration, Oral ; Animals ; Biological Availability ; Coumaric Acids ; administration & dosage ; blood ; pharmacokinetics ; Drugs, Chinese Herbal ; administration & dosage ; analysis ; pharmacokinetics ; Glucosides ; administration & dosage ; blood ; pharmacokinetics ; Iridoids ; administration & dosage ; blood ; pharmacokinetics ; Male ; Rats ; Rats, Sprague-Dawley ; Stilbenes ; administration & dosage ; blood ; pharmacokinetics

In this study, we explored the mechanism of Huganning tablet (HGNP) in the treatment of nonalcoholic fatty liver disease (NAFLD) based on network pharmacology and computer-aided drug design. Firstly, the potential ingredients and targets of HGNP were identified from TCMSP database, Swiss Target Prediction database, Chinese pharmacopoeia (2015) and literatures, and then the targets of HGNP intersected with NAFLD disease targets that obtained in GeneCards database to acquired potential targets. The bioconductor bioinformatics package of R software was used for gene ontology (GO) enrichment and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis. The network of “potential ingredient-key target-pathway” was formed in Cytoscape software to study the interactions between potential ingredients of HGNP, key targets, pathways and NAFLD. Based on the results of network pharmacology, the molecular docking analysis of the key targets and potential active ingredients in HGNP tablets with top degree in the network was conducted using Discovery Studio 2020 software, followed by molecular dynamics simulations, binding free energy calculation, drug-likeness properties analysis and ADMET (absorption, distribution, metabolism, excretion and toxicity) properties prediction. In vitro, HepG2 cells were used to establish steatosis model, and the effects of five key compounds on hepatocyte steatosis were analyzed by oil red O staining and triglyceride (TG) content determination. The results showed that 141 ingredients and 151 potential targets were obtained. A total of 2 526 items and 151 pathways were identified by GO and KEGG enrichment analysis. The molecular docking suggested that five components, isorhamnetin, salvianolic acid B, emodin, resveratrol and rhein, exhibited strong binding ability with key targets [retinoic acid receptor RXR-alpha (RXRA), tumor necrosis factor (TNF), glycogen synthase kinase-3 beta (GSK3B), serine/threonine-protein kinase 1 (AKT1)]. It was further verified that isorhamnetin and salvianolic acid B bind to key targets with good structural stability and binding affinity based on molecular dynamics simulations and binding free energy calculations. The drug-likeness properties, pharmacokinetic properties and toxicity of five key compounds were more comprehensively analyzed through drug-likeness properties analysis and ADMET properties prediction. In vitro, all five compounds, isorhamnetin, salvianolic acid B, emodin, resveratrol, and rhein, improved hepatocyte steatosis of HepG2 cells, confirming the reliability of the present study. In conclusion, based on network pharmacology, computer-aided drug design and in vitro validation, this study investigated the mechanism of HGNP for the treatment of NAFLD at multiple levels and provided a basis for its clinical application.

Objective:To observe the effect of electroacupuncture (EA) at Jiaji (EX-B 2) points plus hydro-acupuncture with sinomenine hydrochloride for low back pain caused by compression fractures in the elderly.Methods:Ninety-five elderly in-patients with low back pain caused by compression fractures were randomly divided into an observation group and an EA group according to the visit sequence.Both groups received the same basic treatment.In the EA group,48 cases were treated with EA at Jiaji (EX-B 2) points plus the basic therapy;47 cases in the observation group received the basic treatment plus EA and hydro-acupuncture with sinomenine hydroch｜oride at Jiaji (EX-B 2) points.The levels of osteoprotegerin (OPG) and interleukin-1β (IL-1β) in peripheral blood were measured by enzyme-linked immunosorbent assay (ELISA) before and at the 21st day of treatment in both groups.Oswestry disability index (ODI) and visual analog scale (VAS) scores were used to analyze the clinical efficacy.Results:After treatment,the OPG content in the observation group was higher with statistical significance compared with that before treatment in the observation group and after the treatment in the EA group,respectively (both P＜0.05);the content of IL-1β,ODI and VAS scores were lower than those before treatment in the observation group and after treatment in the EA group with statistical significances (all P＜0.05).Conclusion:The combination of EA and hydro-acupuncture with sinomenine hydrochloride at Jiaji (EX-B 2) points is effective for low back pain caused by compression fractures in the elderly,and is superior to EA at Jiaji (EX-B 2) points alone.

Objective To investigate the adverse effects of taurine on rabbit corneal endothelial cells. Methods Six rabbits (12 eyes) were selected, and 6 histologic sections were prepared from each of the eyes. Rabbit corneal endothelial cells were cultured by explant culture method. Cells were innoculated on a 12-well tissue culture plate, 2%, 4%, 6%, 8% and 10% taurine solutions were added respectively (cells from the right and left eyes of the same rabbit were added the same concentration of taurine solution), and blank control was established. The growth of corneal endothelial cells was observed by inverted microscopy, and cell morphology on the 1st, 2nd, 4th, 6th and 8th day of culture was observed with Wright staining. Results Corneal endothelial cells cultured with 2%, 4% and 6% taurine solutions and those of blank control formed endothelial cell layers after culture for one week, and the cells exhibited hexagonal or round-like morphology. Corneal endothelial cells cultured with 8% taurine solution appeared to be undergrowth with small cell body on the 4th day, and cell death occurred on the 8th day. Corneal endothelial cells cultured with 10% taurine solution turned out to be undergrowth with small cell body on the 2nd day, and cell death had occurred. The same growth velocity and cell morphology were observed in the corneal endothelial cells from the right and left eyes of the same rabbit. Conclusion Taurine with concentration between 2% and 6% has no adverse effects on the growth of rabbit corneal endothelial cells.

Objective To explore the methods and mid-term results of interventional therapy for atherosclerotic occlusive disease of the superficial femoral artery(SFA).Methods From January 2005 to August 2008,Nineteen cases with 22 diseased lower limbs were admitted.According to Fontaine stage system,there were 6 stage Ⅱb limbs(moderate to severe intermittent claudication),8 stage Ⅲ limbs(rest pain),and 8 stage Ⅳ limbs(6 with ulcer and 2 with gangrene).The mean lesion length was 8.8 cm(5～13 cm).On the basis of the TransAtlantic Inter-Society Consensus(TASC)femoropopliteal classification,the lesions were classified as type B in 4 limbs,type C in 17,and D in 1.Follow up examinations with color duplex ultrasound and/or arteriography were carried out to determine the patency.The improvement of clinical symptoms after operation was analyzed.Results 17 limbs were revascularization successfully,2 limbs were turned to bypass operation,3 limbs were judged untreatable.The technical success rate is 77.3%,and the clinical improvement of symptoms was achieved in 100% of the 17 limbs on which the procedure was successful.The 17 limbs were followed up for a mean of 12.5 months(range 3-33 months).One patient died of cardia infarction one month postoperatively.SFA occlusion happened in 4 limbs.At one year postoperatively,the patency rate was 75%(13/17)after primary operation.Conclusions Primary endovascular treatment of SFA occlusion diseases is a safe,minimally invasive,and effective method.

Objectives To explore the effects of aging on midgastric transverse band(MTB)and gastric emptying.Methods In our prospective study,57 healthy volunteer were divided into young, middle and old-age groups.After taking test meal labeled by 99mTc-iethylenetriaminepentaacetic acid(99mTc-DTPA), the pictures were collected using double probe single photon emission computed tomography(SPECT).Stomach in each frame of the pictures was divided into proximal, midgastric transverse band(MTB)and distal parts.And half gastric emptying time,gastric remnant rate at 90 min, areas of different gastric parts were tested and calculated respectively.Results Half gastric emptying time of whole stomach was(43.24± 11.87)min,(42.07 ± + 9.31)min and(45.81 ± 10.73)min in young,middle and old-age groups, respectively, with P＞0.05.Among young, middle and oldage groups, half gastric emptying time was(38.09 ± 10.26)min,(37.33 ± 9.28)min and(26.74 ± 12.07)min in proximal stomach, and it was(38.35 ± 12.96)min,(37.73 ± 7.46)min and(46.41 ± 10.74)min in distal stomach,respectively.The half gastric emptying time was significantly reduced in proximal stomach and increased in distal stomach in old-age group(both P＜0.05).The gastric nuclide remnant rate at 90min in total stomach was(30.38 ± 19.32)％,(29.03 ± 10.36)％ and(31.92 ± 13.47)％ ,in young, middle and old-age groups, respectively, with P＞0.05.This rate in proximal stomach was(25.01 ± 12.35)％,(26.36± 15.29)％ and(19.54±8.47)％ among three group, respectively.The rate in proximal stomach was(42.25 ± 12.36)％,(41.56 ±± 9.33)％ and(56.05 ± 11.72)％ among three group,respectively.The gastric remnant rate at 90min was reduced in proximal stomach and was increased in distal stomach significantly in old-age group(both P＜0.05).Compared with young and middle-age group,the old-age group showed no difference in areas of total stomach in all the time, while the areas were reduced in proximal stomach and increased in distal stomach significantly from 30 min to 90 min(all P＜0.05).Total stomach versus proximal and distal stomach showed no difference in count/pixel ratio in all time,while a count/pixel ratio was reduced in proximal stomach and increased in distal stomach significantly from 30 min to 90 min(both P＜ 0.05).Both areas and count/pixel ratio of MTB at 60 min and 90 min were significantly increased in old-age group(both P＜0.05).Conclusions The total gastric emptying is not delayed along with aging, while the gastric emptying is increased in proximal stomach and reduced in distal stomach in the elderly.This abnormity of intragastric distribution of food might be related with larger area of midgastric transverse band.

Objective To analyze the distribution and subcellular localization of NOR1 alternative splice isoforms in human tissues and cell lines. Methods NOR1 open reading frame(ORF) was amplified from human fetal brain cDNA library and subcloned into pCMV/myc vector. The level of NOR1 mRNA in human tissues was determined by real-time RT-PCR. Region spanning exon 2 was amplified from cDNA or genomic DNA by specific primers and sequenced. The expression plasmids of NOR1 were transfected into cells and immunofluoresence assay was performed to determine the subcellular localization of NOR1 protein isoforms in human cells. Results The expression of NOR1 mRNA was high in human adult testis, moderate in human fetus nasopharynx, trachea, brain, and kidney tissues, and weak or undetectable in other tissues. Two splice variants of human NOR1 gene resulted from alternative splicing at exon 2 were identified. Both of isoform 1 and isoform 2 mRNA were detected in human fetus brain. Isoform 2 was the sole isoform in other tissues but brain. Only isoform 2 mRNA was detected in cell lines used in this study, though no genomic deletion of exon 2 could be found in all these cell lines. Immunofluoresence assay showed both isoform 1 and isoform 2 proteins were distributed in cytoplasm. Conclusion Alternative splice isoforms of tumor suppressor gene NOR1 are identified. NOR1 isoform 1 and isoform 2 are both detected in fetus brain. NOR1 isoform 2 lacking of exon 2 is the sole isoform in multiple tissues except for brain. The exon 2 encoded peptide does not affect the subcellular location of NOR1 protein.