AIM: The effective antisense sequences targeted VEGF mRNA with computer software would be screened and designed, and effect of them on growth K562 cells and protein expression of VEGF were studied with experiments. METHODS: Seven antisense sequences were selected and synthesized, which consisted of 18-20 deoxynucleotide acid and were modified with phosphorothioate, according to principle of low free energy of overall △G 37 Overall. Cell growth was assayed by trypan blue dye exclusion assay and level of VEGF protien in the media was determined by ELISA. RESULTS: Six of seven sequences were capable of inhibing growth of K562 cells and downregulating the VEGF protein expression significantly, compared with Scrambed control group. It was found that there was a close correlation between low level of overall △G 37 and antisense effectiveness ( r =0 887, P

AIM: To investigate inhibition of K562 cell growth by antisense drug targeted VEGF mRNA. METHODS: X7, 20-mer antisense sequences were selected, synthesized and modified with phosphorothioate. The drug was transfected into K562 cells in the present of lipofection. Cell growth was assayed by trypan blue dye exclusion assay and MTT. The level of VEGF protein in the media was determined by ELISA. The morphology of apoptotic cells were observed by Giemsa staining, and the propotion of apoptotic cells was detected by flow cytometry. RESULTS: The antisense drug inhibited growth of K562 and downregulated expression of VEGF protein significantly, compared with Scrambed control group and showed dose-dependent relation. Signs of apoptosis of K562 cells were not observed. CONCLUSION: Inhibition of K562 cell proliferation, but not cells apoptosis induction is the mechanism of inhibing growth of K562 cells by antisense drug targeted VEGF mRNA. At same time, VEGF has function of promoting K562 cell proliferation, and VEGF mRNA may be a new target attached by drugs. [

Aim To determine the effects of Qiliqian-gxin injected into lateral ventricle on Cardiac function, angiotensin Ⅱ( Ang Ⅱ) , angiotensin converting en-zyme(ACE), angiotensin type 1 receptor(AT1R) and the sympathetic nervous system in the hypothalamic pa-raventricular nucleus of rats with chronic heart failure. Methods Rat model of heart failure was prepared by acute myocardial infarction that was induced by ligation of the left anterior descending coronary artery. Four weeks after heart failure, Qiliqiangxin and Losartan were continuously administered via a syringe pump in-jector connected to lateral ventricle. After four weeks, echocardiogram was used to evaluate the cardiac func-tion and HE was used to observe myocardial tissue morphology, and enzyme linked immunosorbent assay was used to measure plasma norepinephrine( NE) , ser-um NT-proBNP and Ang Ⅱ in the paraventricular nu-cleus. The expression of ACE and AT1 R at mRNA and protein levels in the paraventricular nucleus was deter-mined by Real-time PCR and Western blot, and the RSNA was measured by PowerLab in anesthetized rats. Results Compared with the sham control, the cardiac function was significantly lower while the AngII, ACE, AT1 R expression in the paraventricular nucleus and RSNA were significantly increased in rats with heart failure. Compared with heart failure control, Qiliqian-gxin and Losartan decreased the RSNA and the AngII, ACE, AT1 R expression in the paraventricular nucleus. Conclusion Giving traditional Chinese medicine to the lateral ventricles can decrease the activation of the RAS system, reduce the renal sympathetic nerve activi-ty and improve cardiac function.

Data mining, as known as knowledge discovery in databases, is a non-trivial process of revealing the implied, previously unknown and potentially useful information from the massive data. In recently years, the applications of data mining in the field of pharmaceutical research of traditional Chinese medicine have widespread. Especially in the field of the heritage of experiences of na-tional medical masters, data mining plays an important role. In this study, we would expound of the use of methods of data mining in the heritage of experiences of national medical masters, and analyze their advantages and disadvantages, such as association rules, Bayesian networks, neural networks, and decision trees.
Data Mining ; Databases, Pharmaceutical ; Medicine, Chinese Traditional ; Neural Networks (Computer)

Objective To explore the factors influencing serum trough concentration of vancomycin in pediatric patients with severe gram-positive cocci pneumonia. Methods The general information, the biochemical test results, and plasma concentration of vancomycin were collected from 93 pediatric patients with severe gram-positive cocci pneumonia. The relative factors influencing trough concentration of vancomycin were analyzed retrospectively. Results With the dosage of 40-60 mg/(kg·d), serum trough concentration of vancomycin were between 10-20 mg/L in 26 patients, <10 mg/L in 54 cases, ≥20 mg/L in 13 cases. The ALT, AST, GFR, and γ-GT were significantly different among three groups (P<0.05); the 10-20 mg/L group had the highest levels of AST and γ-GT, the ≥20 mg/L group had the highest level of ALT and the lowest level of GFR. Multiple linear regression analysis showed that GFR was negatively linearly correlated with the serum trough concentration of vancomycin (R2=0.039, P<0.05). The median serum trough concentration of vancomycin in pediatric patients with GFR≥90, 60–90, 30–60 mL/(min·1.73m2) were 8.66, 18.21, 8.45 mg/L respectively, and the difference is statistically significant (P<0.05). Conclusions The serum trough concentration of vancomycin is negatively linearly correlated with GFR in pediatric patients with severe gram-positive cocci pneumonia. The patients with impaired renal function are easier to reach the target serum trough concentration of vancomycin. Clinical use of vancomycin should follow the low doses in the range the guideline recommended, and the serum trough concentration should be closely monitored.

To analyze the composition regularity of prescriptions containing Scutellaria baicalensis in Drug Standard of Ministry of Public Health of the Peoples Republic of China--Chinese Patent Medicines and Preparations on the basis of the traditional Chinese medicine inheritance support system (TCMISS), in order to provide reference for new drug R&D. the platform's software V2.0 was applied to establish a database of prescriptions containing S. baicalensis. The software's statistical statement module, association rules and improved mutual information method and other data mining technologies were adopted to analyze commonly used drugs, combination rules and core combination of S. baicalensis prescriptions. Having analyzed 477 prescriptions containing S. baicalensis, the researchers summarized 45 most commonly used drug combinations, whose ingredients mostly had functions of heat-clearing and damp-drying, purging fire for removing toxin and hemostasis. Drugs adopted in core combinations were relatively concentrated and selected according to definite composition methods. There were 23 diseases that S. baicalensis were most frequently applied in the treatment. Having compared three highly frequent diseases--cold, cough and dizziness, the researchers concluded that S. baicalensis could show different therapeutic effects through different combination ratios. Therefore, TCMISS (V2.0) is an important tool in analyzing the composition regularity of traditional Chinese medicines. The longitudinal and parallel comparison method is an effective method for studying the clinical composition regularity of S. baicalensis, while providing reference for new drug R&D.
China ; Drug Compounding ; statistics & numerical data ; Drug Therapy ; statistics & numerical data ; Humans ; Plant Extracts ; analysis ; therapeutic use ; Scutellaria baicalensis ; chemistry

OBJECTIVETo explore the effects of antisense phosphorothioate oligodeoxynucleotides (AS PS-ODN) of vascular endothelial growth factor (VEGF) on drug-sensitivity of AML and CML cells to homoharringtonine and its possible mechanism.

METHODSA7, which was the most effective AS PS-ODN selected by computer aid-designing and experimental assay, contains 20-mer modified with phosphorothioate. It was transferred into cells by lipofectin while cultured in 2% serum medium for 8 h and then in 10% serum medium. Twenty four hours later, homoharringtonine was added into the culture and cultured for another 48 h. Cell viability was detected by trypan blue exclusion every 24 hours, cell apoptosis by flow cytometry, and level of VEGF protein by a VEGF ELISA kit.

RESULTSThe combination of A7 and homoharringtonine was able to inhibit cell survival (AML cell survival reduced 38%, CML cell survival reduced 25%), down-regulate VEGF protein expression (VEGF protein expression of AML cell and CML cell declined 25.44% and 30.81%, respectively) and increase homoharringtonine induced apoptosis in both AML and CML cells (apoptotic rates of AML and CML cells increased 48.29% and 52.76%, respectively).

CONCLUSIONThe VEGF AS PS-ODN is able to enhance the drug-sensitivity to homoharringtonine, suggesting that inner VEGF proteins in myeloid leukemia cells had a drug-resistant effect.

Acute Disease ; Adult ; Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; genetics ; physiology ; Cell Survival ; drug effects ; genetics ; physiology ; Drug Screening Assays, Antitumor ; methods ; Female ; Harringtonines ; pharmacology ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; pathology ; Leukemia, Myeloid ; genetics ; pathology ; Male ; Middle Aged ; Oligonucleotides, Antisense ; genetics ; Transfection ; methods ; Tumor Cells, Cultured ; Vascular Endothelial Growth Factor A ; genetics ; metabolism ; physiology ; Young Adult

Aim To observe the effect of“Ying-nutrient and Wei-defense unblocking collaterals prescription”on the carotid sympathetic nerve in early atherosclero-sis. Methods The rabbits were randomly divided into 6 groups:Control group, high-fat group, Complex mod-el group, GSTL-H group, GSTL-L group and ATO group. The control group was fed with common food-stuffs, High-fat group with high-fat diet to build early atherosclerosis model, and all the other groups with high-fat diet combined with carotid artery cannula to build early atherosclerosis carotid artery injury rabbit model. All groups were given corresponding medicines intragastrically once a day for 4 weeks. The GSTL-H and GSTL-L group was given Guishaotongluo ultrafine powder 0. 6 g·kg-1·d-1, 0. 3 g·kg-1·d-1, and the ATO group was given suspension of atorvastain so-lution 2. 5 mg · kg-1 · d-1 . Biochemical method was used to detect blood lipid change. HE staining was ap-plied to observe the pathological morphology of intima-media, The content of NE in the carotid arterial was detected by ELISA. The immunohistochemical staining was used to detect the protein expression of NGF, TH around the carotid arterial. The expression of NGF, TH in the carotid artery adventitia was detected by Western blot. Results Compared with normal group, the levels of TC,TG and LDL-C in high-fat group and complex model group were significantly increased( P<0. 01). The degree of the intimal hyperplasia,the con-tent of NE and sympathetic density ( NGF, TH protein expression and distribution ) of the cartoid artery in complex model group were heavier compared with those in high-fat group; the lipid levels, degree of the inti-mal hyperplasia and sympathetic density ( NGF, TH protein expression and distribution ) in the GSTL-H, GSTL-L group were milder in varying degrees compa-ring with those in the complex model group(P<0. 05, P<0. 01 ) . Conclusion “Ying-nutrient and Wei-de-fense unblocking collaterals prescription” can reduce rabbit carotid atherosclerosis, and the mechanism may be related to regulating the sympathetic nerve of arterial wall.

OBJECTIVETo explore action mechanism of electroacupuncture (EA) at "Fenglong" (ST 40) for treatment of hyperlipidemia.

METHODSForty SPF-grade SD rats were randomly divided into a normal group (group A), a model group (group B), a model diet-control group (group C), a model + EA group (group D) and model diet-control + EA group (group E), 8 cases in each one. The rats in group A were fed with normal diet continuously while those in the rest 4 groups were fed with high-fat diet to establish hyperlipidemia model. Afterwards, the rats in group C and group E were fed with normal diet, while EA at "Fenglong" (ST 40) was applied in group D and group E, 30 min per time, once a day. After 28 days of treatment, macrophage was collected in each group. Oil red O-staining was applied to detect the formation of foam cells, and enzyme-linked immunosorbent assay (ELISE) was adopted to measure the contents of total cholesterol (TC) in macrophage and analyze the rate of cholesterol efflux.

RESULTSThe counts of positive cells of oil red O-staining and the contents of TC in the group B, group C and group D were significantly increased compared with those in the group A (all P < 0.01). The counts of positive cells and contents of TC in the group C, group D and group E were significantly clecreased compared with those in the group B (all P < 0.01), and the decline in group D was more obvious than that in the group C (all P < 0.01). Compared with group C and group D, the counts of positive cells and contents of TC in the group E was obviously decreased (all P < 0.01), which was not statistically different from group A (P > 0.05). In the meantime, compared with group C, the rate of cholesterol efflux in group D and group E was significantly increased (both P < 0.01), and the rise in group E was more obvious than that in the group D.

CONCLUSIONThe electroacupuncture at "Fenglong" (ST 40) could significantly prohibit the transformation of macrophage into foam cell and increase rate of cholesterol efflux in macrophage, which could prevent and reverse the formation of foam cell and play an essential role in treating hyperlipidemia and stopping it from developing into a further level.

Acupuncture Points ; Animals ; Biological Transport ; Cells, Cultured ; Cholesterol ; metabolism ; Electroacupuncture ; Foam Cells ; metabolism ; Humans ; Hyperlipidemias ; metabolism ; therapy ; Macrophages ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley

[ ABSTRACT] AIM:To investigate the transient outward potassium channel protein expression in paraventricular nucleus (PVN) and its contribution to renal sympathetic nerve activity (RSNA) in rats with chronic heart failure (CHF). METHODS:A rat model of CHF was prepared by acute myocardial infarction that was induced by ligation of the left ante -rior descending coronary artery .Four weeks after heart failure , echocardiogram was applied to identify the CHF model and plasma norepinephrine (NE), serum NH2-terminal pro-brain natriuretic peptide (NT-proBNP) were detected by ELISA. The expression of ransient outward potassium channel proteins Kv 4.2 and Kv4.3 at mRNA and protein levels was deter-mined by real-time PCR and Western blot .The mean arterial pressure ( MAP) , heart rate ( HR) and RSNA were measured in anesthetized rats with PVN microinjection of potassium channel blockers 4-AP.RESULTS:In CHF group , the rat car-diac function and Kv4.2 and Kv4.3 expression in PVN were obviously lower while plasma NE and serum NT-proBNP were obviously higher than those in sham group .Microinjection of 4-AP into PVN induced an increase in MAP , HR and RSNA in both sham and CHF rats , while the CHF rats exhibited smaller responses to 4-AP than sham-operated rats .CONCLU-SION:Downregulation of Kv4.2 and Kv4.3 expression in the PVN may be a potential mechanism for sympathoexciation in the rats with chronic heart failure .