OBJECTIVETo develop a best chitosan film for using as a drug sustained-release system through the evaluation of the sustained-release property, degradation property, and cytotoxicity to osteoblast.

METHODSOrthogonal experiments were designed to determine the best combination of chitosan film preparations. Drug release rate was determined with Coomassie brilliant blue G250. In a separate study, chitosan films were placed into the test tubes with buffer solution and 10(7) U/L lysozyme. The degradation rate was calculated. Osteoblasts derived from fetal rat calvarial were cultured on chitosan films. Cell proliferation was tested by methyl thiazolyl tetrazolium (MTT) assay. The relative growth rate was calculated and the cytotoxicity was graded.

RESULTSThe best processing condition was 1% acetic acid, chitosan concentration of 2 mg/mL, 6% sodium tripolyphosphate (STPP) concentration, and cross-linking time of one hour. The resulting chitosan film released 33.13% of bovine serum albumin (BSA) within 8 d, 36.73% of BSA within four weeks and the cytotoxicity grade was 0 or 1.

CONCLUSIONThis chitosan film possesses good sustained release property, and a good degradation rate.

Animals ; Chitosan ; Delayed-Action Preparations ; Polyphosphates ; Rats ; Serum Albumin, Bovine

UNLABELLEDOBJECTIVE To study the effects of TiO2 blasted and acid-etched surfaces of cp-titanium on changing composition of oxide-film and attachment and proliferation of osteoblasts in vitro.

METHODScp-titanium discs were prepared and divided into 4 groups: TiO2 blasted (SB), sandBlasted and acid-etching (SLA1 and SLA2) and machine-polished surface (S1). Scanning electron microscopy (SEM), X-ray diffraction (XRD) and electron probe microanalysis (EPMA) were used to test surface morphology and composition of oxide-film. Osteoblasts were cultured on the titanium surface of 4 groups. MTT assay was used to measure the attachment and proliferation.

RESULTSIn regard to surface roughness, average roughness for SB, SLA1 and SLA2 was obviously higher than S0 Oxygen ratio increased on SB,contrarily, it decreased on SLA. A mixture of anatase and rutile-type crystals were observed in the SB. Smaller anatase were observed in the SLA1 and SLA2. The oxide thickness on SLA surface was thiner than that on the SB surface. Alter 1, 4, 24 hours' culture, the number of osteoblast attachment on SB surfaces was the highest (P < 0.05). The number of cells osteoblast proliferation was the highest on SB after 1, 3, 5 and 7 days' culture (P < 0.05).

CONCLUSIONThe thickness and chemical composition of oxide film play an important role in osteoblast attachment and proliferation at the same roughness surface. It is concluded that osteobla.st attachment and proliferation are better on SB surfaces than on SLA1 and SLA2 surfaces.

Cell Adhesion ; Cell Proliferation ; Microscopy, Electron, Scanning ; Osteoblasts ; Oxides ; Surface Properties ; Titanium

Objective To investigate the mechanism of BVT. 2733 on insulin resistance, by using diet-induced obese (DIO) mice model. Methods After having been balanced for 3 days, the C57BL/6J mice were randomly divided into a normal diet group and a high-fat diet (HFD) group. After 20 weeks, the obese mice were further randomly divided into an obese control group, a BVT. 2733 group and a pioglltazone (PGZ) group and they were orally administered with placebo, BVT. 2733 and PGZ separately for two weeks.Adiponectin and leptin mRNA expression levels from adipose tissue were analyzed with real-time quantitative PCR. The levels of plasma glucose, serum insulin and adiponectin were measured with biochemical technology, radioimmunoassay and ELISA. Adipocyte sizes were observed with immunohistocbemistry.Results The body weight, plasma glucose and serum insulin levels raised(P<0.05)in the HFD group and the adipocyte sizes were bigger. Serum insulin levels significantly reduced (P<0.05) and adipocyte sizes reduced, while plasma adiponectin level raised (P<0.01)in the two treatment groups as compared with those in obese controls. Both the mRNA expressions of adiponectin and leptin upregulated(P<0.05)in the PGZ group, but their expressions in the BVT. 2733 group did not alter significantly. The body weight of the mice reduced significantly in the BVT. 2733 group. Conclusion BVT. 2733 can reduce body weight significantly and improve insulin resistance, but cannot influence the expression of adipocytokines.

Taking guizhi fuling capsule (GZFL) for instance, a new method about reference Chinese medicine preparation which was used as standard substance for the quality evaluation of complex Chinese medicine preparation by the fingerprint of reference preparation instead of standard fingerprint was proposed. It could eliminate the errors from different instruments, chromatographic columns and solve the problem of similarity matching in the absence of standard fingerprint. The qualification of reference GZFL was evaluated according to the quality control method of GZFL from Chinese Pharmacopoeia. Then multiple batches of GZFL were estimated, taking fingerprint of reference preparation and standard fingerprint as references, respectively, at different instruments and chromatographic columns. Finally, the packaging and expiration date for reference GZFL were confirmed according to the results of stability investigation. The results indicated that the fingerprint of reference GZFL could be used to assess the quality of GZFL better than standard fingerprint. The data of accelerated stability and long-term stability test demonstrated that reference GZFL was stable in the conditions of double blister package. Therefore, reference GZFL can be used as standard substance in quality control of GZFL.
Capsules ; Drug Stability ; Drugs, Chinese Herbal ; chemistry ; standards ; Reference Standards

OBJECTIVETo study the effects of surface roughness and titanium dioxide (TiO2) layers on commercially pure titanium (cp-Ti) substrates on attachment of osteoblasts in vitro.

METHODS250 pure titanium slices were divided into five groups. Osteoblasts were cultured on five cp-Ti substrates of ground, which blasted with 108-130 microm (S1), 216-301 microm (S2), 356-411 microm (S3) TiO2 particles and titanium-sprayed plasma (TPS) surfaces, surfaces prepared by hand grinding with SiC paper to 600 grits served as control (S0). Surface average roughness and the TiO2 film structure was evaluated. For morphology and attachment measurement, osteoblasts were cultured for 1, 4, 12 and 24 h, evaluated by scanning electronic microscope (SEM) observation and MTr assay.

RESULTSOsteoblasts spread well on the titanium surfaces. Further more, osteoblasts spread more well on S3 surfaces. After 1 and 4 h culture, the number of cells on S3 surfaces was the highest (P < 0.05). The number of cells on S3 surfaces was the same (P > 0.05) as TPS surfaces and higher than other groups (P < 0.05) after 12 and 24 h. The number of cells of all experimental groups were higher than S0 surfaces after 4, 12 and 24 h (P < 0.05).

CONCLUSIONIt was concluded that the coarse TiO2 particles blasted surface would optimize initial osteoblast responses.

Cell Differentiation ; Microscopy, Electron, Scanning ; Osteoblasts ; Surface Properties ; Titanium

OBJECTIVEThe aim of this study was to investigate the mechanism and efficacy of tetramethylpyrazine-eluting stents (TES) on inhibiting neointima formation in porcine coronary arteries.

METHODSTES was prepared by tetramethylpyrazine spray-coated in bare metal stents (BMS). Pigs were implanted with TES or BMS (n = 7 each), respectively. Quantitative coronary angiography (QCA) and intravascular ultrasound (IVUS) were performed before, immediately after stenting and at 28 days after stenting. Coronary arteries segments (5 cm) before and post stenting area (5 cm) as well as at stenting location were harvested at 28 days post stenting for histopathological examinations (inflammation, vascular smooth muscle cells proliferation and apoptosis).

RESULTSFollow up QCA at 28 days showed that percentage diameter stenosis were significantly lower in the TES group than that in the BMS group [(10.0 +/- 2.1)% vs (60.2 +/- 23.5)%, P = 0.01]. The lumen area determined by IVUS was similar between the two groups and there was no in-stent thrombosis in TES or BMS treated animals. Internal elastic lamina area was significantly larger while the neointimal area [(1.51 +/- 0.45) mm(2) vs (4.60 +/- 1.39) mm(2), P = 0.04] was significantly smaller in the TES group than that in the BMS group. Histopathological assessments showed fewer inflammatory cells in the stented-coronary artery walls than those at the border zones of stenting in both groups. The number of proliferating cells were significantly decreased while apoptotic cells were significantly increased in the TES group compared with the BMS group (all P < 0.05).

CONCLUSIONTES could effectively reduce in-stent restenosis in this porcine model by attenuating vascular smooth muscle proliferation and enhancing vascular smooth muscle apoptosis post stenting.

Animals ; Coronary Restenosis ; prevention & control ; Coronary Vessels ; pathology ; Disease Models, Animal ; Double-Blind Method ; Drug-Eluting Stents ; Pyrazines ; administration & dosage ; Swine ; Tunica Intima ; drug effects ; pathology

UNLABELLEDOBJECTIVE To evaluate the effects of grooved, alkali- and heat-treated, acid-etched and TiO2 blasted surfaces of titanium substrates on F-actin cytoskeleton of osteoblasts in vitro.

METHODSOsteoblasts derived from fetal rat calvarial were cultured on 6 different commercially pure titanium discs-grooved(G), sandblasted (SB), sand-blasted and acid-etching (SLA) surfaces and alkali- and heat-treated (AH1, AH2, AH3) surfaces. For F-actin cytoskeleton measurement, osteoblasts whose filamentous actin was stained with phalloidin-TRITC were cultured for 1, 2, 4, 12 h, evaluated by CLSM observation.

RESULTSOsteoblasts attached to the different types of surfaces after 1 hour culture were similar. The actin cytoskeleton formed a ring of cortical filaments around the nucleus after 1 hour on SB, AH2, AH3, SLA surfaces. Actin filaments condensed along edges of pits. The actin filaments of seeded cells were spread after 2 h. The actin filaments on G formed bundles around the nucleus. The filaments began to parallel to the grooves. On AH1, the fibres formed a ring of cortical filaments around the nucleus with some cytoplasmic fibres radially oriented. On AH2, AH3, SB, the fibres orignised in a cytoplasmic meshwork with fibres which terminate at the ridge of depressions. The cell were suspending itself over the depressed areas. Actin filaments on SB were distinct and well formed that were oriented paralled to one another and the long axis of cells. After 4 h, actin filaments appeared organised in a parallel to one another and the long axis of cells. After 12 h, the actin filaments on all surfaces were well spread and were oriented paralled to another and to the long axis of the cell. The filaments formed bundles which reached to holes or adhered to the ridge of raised points, suspending cells over depressed areas.

CONCLUSIONAfter 12 h, the actin filaments on all surfaces were well spread and were oriented parallel to another and to the long axis of the cell. It was concluded that F-actin cytoskeleton of osteoblasts were spread best on SB surfaces among all surfaces.

Actin Cytoskeleton ; Actins ; Animals ; Cytoskeleton ; Microtubules ; Osteoblasts ; Prostheses and Implants ; Rats ; Surface Properties ; Titanium

OBJECTIVETo investigate the distribution of hepatitis B virus (HBV) genotype in Guizhou and to study the relationship between the genotype and the progression of liver disease.

METHODS786 patients with chronic HBV infection, from 4 cities of Guizhou, including 346 asymptomatic carriers (ASC), 313 chronic hepatitis (CH), 77 liver cirrhosis (LC), 50 hepatocellular carcinoma (HCC) were examined. HBV genotype was determined by restriction fragment length polymorphism analysis and the subtypes were determined by direct sequencing of PCR product in 94 patients with HBV B genotype, the relationship between HBV genotype and the progression of liver disease was studied by multifactor analysis such as HBeAg positivity, HBV DNA load and ALT level.

RESULTSOf the 786 patients, 7 (0.89%), 497 (63.23%), 275 (34.99%), and 7 (0.89%) belonged to genotype A, B, C, D, respectively. There was statistically significant difference in the distribution of genotype B among Kaili (96.04%), Zunyi (78.79%), Duyun (64.52%) and Guiyang (53.14%) (P< 0.01). Genotype C was more prevalent in Guiyang than in other three cities (P < 0.01, or P < 0.05). Out of 94 genotypes B, 93 (98.94%) belonged to subtype Ba, only one was subtype Bj. There were statistically significant difference in the distribution of genotype B and C among various stage of liver disease (P < 0.05 or P < 0.01). Genotype B showed a gradual decrease from ASC, CH, LC to the HCC group while in contrast, genotype C showed a gradual increase in the same order. The ALT levels and the mean age were significantly higher and older in patients with genotype C than those in genotype B (P < 0.01 or 0.05). The HBeAg positivity was significantly lower in genotype C than that in genotype B (P < 0.025).

CONCLUSIONData showed that there were genotype A, B, C and D existing in Guizhou. Genotype B was the major one but genotype C was more commonly seen. In genotype B, subtype Ba appeared to be predominant. The geographic distribution of genotype B and C were different in some cities of Guizhou. Compared to genotype B, genotype C was associated with the development of more severe liver damage.

Carcinoma, Hepatocellular ; pathology ; virology ; DNA, Viral ; analysis ; Disease Progression ; Genotype ; Hepatitis B virus ; classification ; genetics ; Hepatitis B, Chronic ; genetics ; pathology ; Humans ; Liver ; pathology ; Liver Cirrhosis ; pathology ; virology ; Liver Neoplasms ; pathology ; virology ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length

To study the pharmacokinetics, excretion characteristics and plasma protein binding rate of asperosaponin VI (A-VI) and its active metabolite hederagenin (M1). A-VI and M1 concentrations in plasma, bile, urine and feces were determined by established LC-MS/MS to calculate the pharmacokinetic parameters. The plasma protein binding rate of A-VI was determined by equilibrium dialysis method. the double peaks was observed in the A-VI plasma concentration-time curve, after rats were orally administered with low, medium and high doses of A-VI (0.03, 0.09, 0.27 g x kg(-1)). The Cmax1 and Cmax2 of A-VI were (28.88 +/- 49.78) and (4.480 +/- 1.872) microg x L(-1), (35.19 +/- 23.53) and (22.11 +/- 16.15) microg x L(-1), (73.37 +/- 37.28) and (132.2 +/- 160.7) microg x L(-1), respectively. The AUC0-t, of A-VI were (43.21 +/- 37.32), (133.9 +/- 102.5) and (779.6 +/- 876.9) microg x h x L(-1), respectively. The t1/2 of A-VI were (3.3 +/- 0.8), (3.2 +/- 2.3) and (4.5 +/- 1.2) h, respectively. The Cmax of M1 were (16.03 +/- 9.336), (26.41 +/- 11.95) and (28.71 +/- 5.874) microg x L(-1), respectively. The AUC0-t, of M1 were (105.6 +/- 73.60), (260.0 +/-153.9) and (323.1 +/- 107.9) microg x h x L(-1), respectively. The t1/2 of M1 were (4.1 +/- 3.4), (4.4 +/- 2.3), (3.9 +/- 0.9) h, respectively. No significant gender difference was found in the in vivo pharmacokinetics of A-VI and M1. There was no accumulation of A-VI and M1 after multiple administrations of A-VI (0.09 g x kg(-1)). After oral administration of A-VI, the double peaks were also observed in biliary and urinary excretion rate-time curves for A-VI. M1 was detected in the feces samples at 6 h after oral administration. The average plasma protein binding rate of A-VI was 92. 9% in rats.
Administration, Oral ; Animals ; Area Under Curve ; Bile ; metabolism ; Drugs, Chinese Herbal ; metabolism ; pharmacokinetics ; Female ; Male ; Plants, Medicinal ; Protein Binding ; drug effects ; Rats ; Saponins ; blood ; metabolism ; pharmacokinetics ; urine

Adult ; Erythema ; etiology ; Female ; Humans ; Leprosy ; complications ; Male