OBJECTIVETo construct a recombinant adenovirus vector containing bone morphogenic protein-7 (BMP-7) gene and to identify its biological activities in proximal tubule epithelial cells (PTECs).

METHODSForty-six fragments of BMP-7 gene were obtained by PCR method and then were ligated to the full-length gene. The full length sequence of BMP-7 was subcloned into pBluescript II(+) vectors, and confirmed by sequencing. Double digested with Not I and Hind III, BMP-7 gene was inserted into pShuttle-CMV. EcoR I pre-linearized pShuttle plasmid was transformed into competence bacterium BJ5183 to obtain the recombinant adenovirus-BMP-7 by efficient homologous recombination. Then the AdBMP-7 was obtained by packaging Pac I linearized in 293 cells. Adenoviral titer was determined by adenovirus fluorescent detection kit. The protein expression of BMP-7 in PTECs was respectively detected by ELISA and Western blot. RT-PCR method was used for analyzing the alpha-smooth muscle action (alpha-SMA) expression in PTECs, which was treated consecutively with TGF-beta and AdBMP-7.

RESULTSThe recombinant plasmid AdBMP-7 was successfully generated, which increased BMP-7 protein expression levels in PTECs, and down-regulated TGF-beta-induced alpha-SMA expression.

CONCLUSIONThe bioactive recombinant adenovirus AdBMP-7 has been successfully constructed, which may be effective in inhibition of chronic renal fibrosis.

Adenoviridae ; genetics ; metabolism ; Animals ; Base Sequence ; Bone Morphogenetic Protein 7 ; biosynthesis ; genetics ; Cells, Cultured ; Epithelial Cells ; metabolism ; Genetic Vectors ; genetics ; Kidney Tubules, Proximal ; cytology ; metabolism ; Molecular Sequence Data ; Rats ; Recombinant Proteins ; biosynthesis ; genetics

Objective: To optimize the forming process of gel patch and explore the compatibility-synergistic mechanism of Aconiti Radix-Zanthoxyli Pericarpium volatile oil gel patch. Methods: According to the indexes of skin followability, disclosing ability, initial adhesion and peeling degree, the substrate-property of eight commercially gel patches was evaluated. Then, with the comprehensive sensory evaluation, initial adhesion, stickiness and peeling degree as the index, single factor experiment and mixture design experiment were used to screen the proportion of matrix excipients of gel patch. Taking the number of acetic acid-induced writhing in mice as the index, the drug loading of gel paste was screened by in vivo animal evaluation. In addition, the compatibility effect of Aconiti Radix-Zanthoxyli Pericarpium volatile oil gel patch was studied by using formaldehyde-induced pain model and metabolomics. Results: Different brands of gel plaster had great difference in their skin followability, disclosing ability, initial adhesion and peeling degree. The initial adhesion and peel degree of gel patch had great influence on its disclosing ability and the greater the initial adhesion and peel degree were, the worse disclosing ability was. The matrix of the Aconiti Radix gel patch was 3% polyacrylic acid sodium NP700, 1.5% carboxymethyl cellulose sodium, 0.25% Carbomer, 0.06% crystalline aluminum chloride, 0.1% ethylenediamine tetra acetic acid two sodium, 26% glycerol, 61.5% water, 4% diatomaceous earth, 0.15% tartaric acid; Drug dosage of each gel patch was 560 mg extracts of Aconiti Radix. Zanthoxyli Pericarpium olatile oil could enhance the analgesic effect of Aconiti Radix gel patch, which might be mainly through glycerol phospholipid metabolism and linoleic acid metabolism. Conclusion: To ensure better adherence and disclosure of gel patch, the initial adhesive force of gel patch should be controlled between 7-18 balls and the peel degree should be controlled between 0.03-0.97 kN/m. The Prepared Aconiti Radix gel patch has a good appearance, adhesion and shape with a clear analgesic effect, and Zanthoxyli Pericarpium olatile oil can enhance analgesic effect of Aconiti Radix gel patch. This study provides a basis for the development and application of new preparations for the external use in Aconiti Radix.

OBJECTIVETaking Chinese compound medicine Yuchangning as a research model, prepared the pH and time dependent Yuchangning tablets for colon-specific delivery (PT-YT-CSD), and evaluated the releasing property in vitro.

METHODThe coating prescription is filtered by the release extent of matrine and oxymatrine in vitro and the wicking rate of the tablet, which including the category and proportion of film forming agent and porogen, the sort and dosage of fluidizing agent, the increment of weight after coating and so on. The releasing property of the preparation is evaluated by the dissolution tests in vitro through measuring the content of matrine and oxymatrine content.

RESULTThe preparation method of the PYTCSD: After prepared plain tablets, the 95% alcoholic solution of EC and Eudragit II are mixed in a 7:3 EC: Eudragit II ratio and then added in DEP up to 10% of the coating amount, reduced the alcohol concentration to 4% by diluting with ethonal. Tablet was coated by the alcohol solution and the weight of the plain tablet was increased by 3%. The dissolution tests in vitro indicated that matrine and oxymatrine were not dissolved in the simulated gastric juice after 2 h. The accumulative quantities of matrine and oxymatrine were less than 10% in the simulated intestinal juice after 4 h. The quantities of matrine and oxymatrine are 75.7% and 76.8% in the simulated colon juice after 1 h.

CONCLUSIONThe PYTCSD was prepared and the preparation could fulfil the aim of delivering in the specific colon in vitro.

Alkaloids ; analysis ; Astragalus membranaceus ; chemistry ; Cellulose ; analogs & derivatives ; Colon ; metabolism ; Delayed-Action Preparations ; Drug Combinations ; Drug Compounding ; Drug Delivery Systems ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacokinetics ; Hydrogen-Ion Concentration ; Intestinal Absorption ; Plants, Medicinal ; chemistry ; Polymethacrylic Acids ; Quinolizines ; analysis ; Sophora ; chemistry ; Tablets, Enteric-Coated

OBJECTIVETo discuss the value of endoscopic technique in maxillofacial and neck regions.

METHODSThis retrospective review included 102 clinical patients with superficial masses in maxillofacial and neck regions. The indications, incisions, endoscopic space preparation and the key points of operative process were analysed.

RESULTAll of the results were satisfying.

CONCLUSIONThe endoscopic technique in treating superficial masses in maxillofacial and neck regions had the advantage of wide indications, multiple incision choices, easy control of operative layers and less complications. And the recent and remote effects were excellent.

Endoscopy ; methods ; Humans ; Neck ; surgery ; Oral Surgical Procedures ; methods ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo investigate the effects of interferon -alpha1b (IFN-alpha1b) on hepatic intercellular adhesion molecule-1 (ICAM-1) expression and serum HBV DNA in patients with chronic hepatitis B.

METHODSBefore and 6 months after IFN-alpha1b treatment, liver biopsy was performed in patients with chronic hepatitis B to detect the expression of ICAM-1 in the liver tissues using immunohistochemistry. Serum HBV load was detected with real-time fluorescence polymerase chain reaction.

RESULTCAM-1 expression in the liver tissue was significantly down-regulated after IFN treatment in patients with severe and moderate chronic hepatitis B (P<0.05). No significant variation was noted in the expression of ICAM-1 in the livers of patients with mild chronic hepatitis B after the treatment (P>0.05). In the patients weakly positive for ICAM-1 expression (+), serum HBV DNA varied scarcely after the treatment (P>0.05), while in the patients with strong ICAM-1 positivity (++, +++, or ++++), significant variation of serum HBV DNA occurred after the treatment (P<0.05 or P<0.01).

CONCLUSIONThe therapeutic effect of IFN-alpha1b is associated with the expression of ICAM-1 in the hepatocytes, and its expression might enhance the effects of IFN on HBV DNA in patients with chronic hepatitis B.

Adolescent ; Adult ; DNA, Viral ; blood ; Female ; Hepatitis B virus ; drug effects ; genetics ; Hepatitis B, Chronic ; blood ; drug therapy ; virology ; Humans ; Immunohistochemistry ; Intercellular Adhesion Molecule-1 ; biosynthesis ; Interferon-alpha ; therapeutic use ; Liver ; drug effects ; metabolism ; virology ; Male ; Middle Aged ; Polymerase Chain Reaction ; methods ; Viral Load ; Young Adult

To increase drug concentration in the head through intranasal administration, we have investigated the excised animal nasal mucosa permeability and nasal toxicity of the baicalin drug carrier systems, such as baicalin liposomes, beta-cyclodextrin inclusion compound, and phospholipid complex. A transport of baicalin drug carrier systems through nasal mucosa was simulated in diffusion chamber in vitro, and swine, caprine and rabbit nasal mucosa was used, the concentration of drug in the receptor was determined by HPLC. By taking the apparent permeability coefficients as evaluation standard, investigated the isolated animal nasal mucosa permeability of different baicalin drug systems was investigated for screening the best baicalin drug carrier system through nasal cavity administration. Toxicity of baicalin and its phospholipids complex on toad palate mucosal cilia movement and rats nasal mucosa long-term toxicity were studied in vivo. The apparent permeability coefficient of three kinds of baicalin drug carrier systems was better than that of baicalin (P < 0.05), and its lag-time was obviously shortened. At the same time, the apparent permeability coefficient of phospholipid complex was higher than those of other two drug carrier systems (P < 0.05). The results showed that the baicalin phospholipids complex nasal mucosa permeability was obviously superior to the other two drug systems. Baicalin phospholipids complex had no toxicity to ciliary movement, and had no irritation to rat nasal mucosa. The results show that baicalin phospholipid complex was the best baicalin drug carrier system, it could significantly enhance the permeability of baicalin across nasal mucosa, had no toxicity to nasal mucosa, and could be used for intranasal administration.
Administration, Intranasal ; Animals ; Bufo bufo ; Drug Carriers ; pharmacokinetics ; toxicity ; Drug Delivery Systems ; Female ; Flavonoids ; administration & dosage ; pharmacokinetics ; toxicity ; Goats ; Liposomes ; pharmacokinetics ; toxicity ; Male ; Nasal Mucosa ; drug effects ; metabolism ; Palate ; drug effects ; Permeability ; Phospholipids ; pharmacokinetics ; toxicity ; Rabbits ; Random Allocation ; Rats ; Swine ; beta-Cyclodextrins ; pharmacokinetics ; toxicity

OBJECTIVETo observe the effects of combined beta(1) adrenergic receptor (AR) antagonist with beta(2)AR agonist therapy on cardiac function and cardiomyocyte apoptosis in heart failure rats.

METHODSHeart failure was induced by isoproterenol and rats were randomly divided into metoprolol group (50 mg/kg twice daily/gavage, n = 11), combined treatment group (fenoterol 125 microg/kg and metoprolol 50 mg/kg twice daily/gavage, n = 11) and placebo group (saline, n = 10), another normal 9 male Wistar rats served as control group. After 8 weeks' treatment, cardiac function, apoptosis index (AI), Caspase-3 activity, expression levels of bcl-2 and bax protein, organ weight/body weight and collagen volume fraction (CVF) were evaluated.

RESULTS(1) Left ventricular end diastolic dimension, left ventricular end systolic dimension and E/A ratio were significantly increased and fractional shortening, ejection fraction significantly reduced post isoproterenol (all P < 0.05 vs. control) and these changes were significantly attenuated by metoprolol alone (all P < 0.05 vs. placebo) and further attenuated by the metoprolol and fenoterol combination therapy (all P < 0.05 vs. placebo and metoprolol). (2) Left ventricular weight to body weight ratio, lung weight to body weight ratio and CVF were also significantly reduced in metoprolol and combined treatment group than those in placebo group (all P < 0.01). (3) Compared with placebo group, AI and Caspase-3 activity were significantly lower in metoprolol group (all P < 0.01 vs. placebo) and further reduced in combined treatment group (all P < 0.01 vs. metoprolol). (4) The expression level of bax protein was significantly lower in metoprolol group while bcl-2/bax significantly higher than those in placebo group. These changes were more significant in combined treatment group (all P < 0.01 vs. metoprolol).

CONCLUSIONSbeta(1)AR antagonist in combination with beta(2)AR agonist further improved the cardiac function and prevented cardiac remodeling compared with using beta(1)AR antagonist alone in heart failure rats. Downregulated bax and upregulated bcl-2/bax expressions might contribute to the observed beneficial therapy effects by reducing cardiomyocyte apoptosis in these animals.

Adrenergic beta-1 Receptor Antagonists ; Adrenergic beta-2 Receptor Agonists ; Adrenergic beta-Agonists ; pharmacology ; therapeutic use ; Adrenergic beta-Antagonists ; pharmacology ; therapeutic use ; Animals ; Apoptosis ; drug effects ; Drug Therapy, Combination ; Heart Failure ; drug therapy ; Male ; Myocytes, Cardiac ; cytology ; Rats ; Rats, Wistar ; Ventricular Remodeling

The aim of the study is to evaluate the effects of the single and mixed decoction of Thallus laminariae (kelp) and Glycyrrhiza glabra (licorice) on the metabolism and their difference. The mixed decoction of kelp and licorice and the single decoction were made and intragastrically administered to the SD rats. The effect on system metabolism, the toxicity of liver and kidney were assessed by GC-MS profiling of the endogenous molecules in serum, routine biochemical assays and histographic inspection of tissues from SD rats, separately. The mixed decoction of kelp and licorice induced more obvious pathological abnormalities in SD rats than a single decoction of kelp, while the extracts of licorice did not show any pathological change. Neither the mixed, nor the single decoction showed abnormal histopathology. After intragastric administration of extracts for 5 days, the mixed decoction induced a decrease of ALT (no significant change in the groups of single decoction) and an increase of BUN (so did the single decoction of kelp). Metabolomic profile of the molecules in serum revealed that the metabolic patterns were all obviously affected for the three groups, i.e., the mixed and single decoction of kelp and licorice. The rats given with the single decoction of kelp showed a similar pattern to that of the mixed decoction, indicating that the kelp primarily contributed the perturbation of metabolism for the mixed decoction. All three groups induced a decrease of branched chain amino acids, TCA cycle intermediates and glycolysis intermediates (e.g., pyruvic acid and lactic acid) and an increase of 3-hydroxybutyric acid. Kelp decoction showed stronger potential in reducing TCA cycle intermediates and glycolysis intermediates than the other two groups, while the levels of branched chain amino acids were the lowest after licorice extracts were given. These results suggested that the effect of the mixed decoction on metabolism was closely associated with both kelp and licorice. The continuous administration of single decoction of kelp and the mixed decoction of licorice and kelp resulted in pathological abnormalities in kidney of SD rats. The mixed decoction of kelp and licorice distinctly perturbed sera molecules and hence system metabolism, which showed associated with those of kelp and licorice. Although the metabolic effect was associated with both kelp and licorice, the results suggested kelp contributed to it primarily.
Animals ; Glycyrrhiza ; chemistry ; Kelp ; chemistry ; Kidney ; drug effects ; Liver ; drug effects ; Metabolomics ; Plant Preparations ; pharmacology ; Rats ; Rats, Sprague-Dawley

BACKGROUNDStimulation of the heart beta 3-adrenoceptor (AR) may result in a negative inotropic effect. Being up-regulated, beta 3-AR plays a more important role in the regulation of cardiac function during heart failure. However, the effect of chronic blocking of beta 3-AR on heart failure has not been fully elucidated. In this study, we used a selective beta 3-AR antagonist SR59230A to treat a well defined heart failure rat model chronically, then evaluated its effect on cardiac function and investigated the mechanism.

METHODSMale Wistar rats were chosen randomly as controls (n = 8). Isoproterenol induced heart failure rats were randomly divided into ISO group (n = 10) and SR group (n = 10). The ISO group received intraperitoneal injection of 1 ml saline twice a day; the SR group received intraperitoneal injection of SR59230A 85 nmol in 1 ml saline twice a day; and the control group received no treatment. The treatment was started 24 hours after the last isoproterenol injection and continued for 7 weeks. Then we measured the following indexes: the ratio of heart weight to body weight (HW/BW) and the ratio of left ventricular weight to body weight (LVW/BW), collagen volume fraction (CVF), left ventricular end diastolic dimension (LVEDd), left ventricular end systolic dimension (LVESd), ejection fraction (EF), fractional shortening (FS) and the ratio of E wave to A wave (E/A), the mRNA and protein expression of beta 3-AR and eNOS, and cGMP level in the heart.

RESULTSThe ratios HW/BW and LVW/BW were significantly increased in the ISO group compared with the control group (P < 0.01), but they were limited in the SR group (P < 0.05 compared with the ISO group). CVF increased in the ISO group and the SR group (P < 0.01), but it was significantly attenuated in the SR group (P < 0.01). LVEDd, LVESd and E/A ratio were significantly increased in the ISO group compared with the control group (P < 0.01), while EF and FS were significantly decreased (P < 0.01). Compared with the ISO group, the SR group showed that LVEDd, LVESd and E/A ratio were significantly decreased (P < 0.01), whereas EF and FS were significantly increased (P < 0.01). beta(3)-AR and eNOS mRNA and protein in the ISO group were significantly increased when compared with the control group (P < 0.01). These increases were all attenuated in the SR group compared with the ISO group (P < 0.01). The level of cGMP in myocardial tissue was significantly increased in the ISO group compared with the control group (P < 0.01), whereas SR59230A treatment normalized this increment (P < 0.01).

CONCLUSIONSChronic blocking of beta 3-AR could ameliorate cardiac function in heart failure rats and its mechanism involves inhibition of the negative inotropic effect and attenuation of cardiac remodeling.

Adrenergic beta-3 Receptor Antagonists ; Adrenergic beta-Antagonists ; pharmacology ; therapeutic use ; Animals ; Blotting, Western ; Disease Models, Animal ; Echocardiography ; Enzyme-Linked Immunosorbent Assay ; Heart Failure ; drug therapy ; physiopathology ; Male ; Myocardium ; pathology ; Nitric Oxide Synthase Type III ; genetics ; Propanolamines ; pharmacology ; Rats ; Rats, Wistar ; Receptors, Adrenergic, beta-3 ; physiology ; Reverse Transcriptase Polymerase Chain Reaction ; Ventricular Function, Left ; drug effects

OBJECTIVETo determine the reference value of plasma N-terminal pro-B-type natriuretic peptide (NT-proBNP) in subjects without heart diseases.

METHODSThe plasma concentration of NT-proBNP was measured with ELISA method in 300 adults excluded heart disease through various examinations including electrocardiography, echocardiography, X-ray and coronary artery angiography. The plasma NT-proBNP concentration was compared between age-groups 30-39, 40-49, 50-59, 60-69 and > or = 70 years old, between male and female in the same age-group and between subjects with and without hypertension, diabetes and obesity. A multiple linear regression analysis was used to detect factors influencing NT-proBNP among age, sex, body mass index, blood pressure, heart rate, serum creatinine, hypertension, diabetes mellitus, use of angiotensin-converting-enzyme inhibitors, Ca(2+)-antagonist, and beta-blocker.

RESULTSThe plasma NT-proBNP concentration increased in proportion to aging in male subjects more than 60 years old (P < 0.05), remained unchanged in males less than 60 years old and females (P > 0.05). Plasma NT-proBNP concentration was significantly higher in female (170-660 pmol/L) than in male (160-470 pmol/L) in subjects less than 60 years old (P < 0.05) and significantly lower in female (180-560 pmol/L) than in male (180-760 pmol/L) in subjects more than 60 years old (P < 0.05). Multiple linear regression analysis demonstrated that age was the only independent predictor for plasma NT-proBNP in these subjects (P < 0.01).

CONCLUSIONThe plasma concentration of NT-proBNP in subjects without heart diseases was different between male and female, and was increasing with age in male subjects more than 60 years old.

Adult ; Age Factors ; Aged ; Chest Pain ; blood ; Female ; Humans ; Linear Models ; Male ; Middle Aged ; Natriuretic Peptide, Brain ; blood ; Peptide Fragments ; blood ; Reference Values