AIMTo determine clonidine in rabbit plasma by LC-MS.

METHODSThe LC-MS system consisted of Waters Alliance 2790 HPLC and Micromass ZQ-4000 MS. The HPLC was performed by using XTerra C18 (150 mm x 2.1 mm ID, 5 microm). The mobile phase, consisting of acetonitrile/ammonium hydrogen carbonate solution, was maintained to a flow-rate of 0.2 mL x min(-1) and the linear gradient elution was adopted. Mass spectrum was obtained by using electrospray ionization interface and the m/z of SIM was 230.

RESULTSThe average recovery was high and the method was reproducible. The calibration curve showed good linearity in the range of 1 - 80 microg x L(-1), the lowest limit of detection was 0.05 microg x L(-1). The Cmax, AUC0-t, and Tmax value of the pharmacokinetics parameter were (27 +/- 9) microg x L(-1), (5,352 +/- 1,121) microg x L(-1), (79 +/- 17) h.

CONCLUSIONThe results demonstrated that the method had high sensitivity, good selectivity, accuracy and precision. It is used to determine the clonidine concentration in plasma. The transdermal patch can deliver clonidine to the surface of rabbit skin stably for periods of up to 1 week after a single application.

Administration, Cutaneous ; Animals ; Antihypertensive Agents ; administration & dosage ; blood ; pharmacokinetics ; Area Under Curve ; Chromatography, High Pressure Liquid ; methods ; Clonidine ; administration & dosage ; blood ; pharmacokinetics ; Rabbits ; Spectrometry, Mass, Electrospray Ionization ; methods

AIMTo study the effect of lidocaine-dodecanol binary eutectic system on the transdermal permeation of lidocaine.

METHODSBinary eutectic mixture of different proportions of lidocaine and dodecanol were prepared and the patch containing the binary eutectic mixture was developed. The solubilities of pure lidocaine and lidocaine from the binary eutectic system were determined in pH 7.9 phosphate buffer. The transdermal flux of lidocaine from the patches containing the binary eutectic system and pure lidocaine were measured using Franz-type single diffusion cell.

RESULTSThe melting point of the lidocaine-dodecanol binary eutectic system was markedly lower than that of pure lidocaine. The steady state transdermal flux of lidocaine from the patch of the binary eutectic system was six times as much as that of pure lidocaine patch.

CONCLUSIONThe lidocaine-dodecanol binary eutectic system could produce high thermodynamic activity of the drug and the high driving force for transdermal permeation of lidocaine.

Administration, Cutaneous ; Anesthetics, Local ; administration & dosage ; pharmacokinetics ; Animals ; Dodecanol ; administration & dosage ; chemistry ; Drug Combinations ; Drug Stability ; Guinea Pigs ; Lidocaine ; administration & dosage ; pharmacokinetics ; Skin Absorption ; Solubility

OBJECTIVEThis experiment is to test the survival enhance effect of leptin in grainy fat transplantation.

METHODSThe 0.6 g grainy fat was obtained from the Wistar mouse subcutaneous tissue in the abdomen. It was then mixed with 0.9% NS and 20 ng/ml leptin and transplanted in the head subcutaneously. The transplanted tissue was taken out after 10, 20 and 40 days. After being weighed, the tissue was dyed with HE staining and VEGF staining. The data was analyzed with SPSS software.

RESULTSThe fibrous membrane around the tissue in leptin group was thin and there was less fat cell necrosis, compared with the control group. The remained-weight rate was 108.3%, 83.3% and 66.3% in 10, 20 and 40 days after the surgery. The differences were statistic significant between the leptin groups and the control (P < 0.05). The examinations of the HE and VEGF stain showed that there was the rich vascularization in leptin treated tissue.

CONCLUSIONLeptin could enhance the survival of the transplanted grainy fat due to its angiogenesis effect.

Adipose Tissue ; transplantation ; Animals ; Female ; Humans ; Leptin ; pharmacology ; Male ; Models, Animal ; Neovascularization, Physiologic ; drug effects ; Rats ; Rats, Wistar ; Survival Analysis ; Time Factors ; Transplantation, Homologous ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo determine the optimal drill area on the footplate with the 3D measurements of the stapes and the vestibular end organs.

METHODSFour temporal bones were extracted from the fresh cadavers and undecalcified polymer-embedded. After serially sectioning, image processing and the 3D precisely reconstruction, a local Cartesian coordinates was established in which the tympanic surface of the footplate was supposed to be XY plane and the Z coordinate axis passed through the central point of the footplate and was vertical to the XY plane. The configurations of the utricle and saccule were delineated quantitatively, and then any distance between one point on the surface of the footplate and another point on the surface of the utricle or saccule and its orientation can be measured.

RESULTSThere was a "V" shaped cleft between the utricle and the saccule. The angle of the" V" shaped cleft was 50.31 +/- 19.90 (17.00 - 68.00) degrees. The apex of the cleft directed anterosuperiorly and approached the footplate center, while beneath the posteroinferior part of the footplate was an open and deep area. The vertical distance from the center point of the footplate to the vestibular end organs was (2.20 +/- 0.548) mm, the maximum of 3.0 mm and the minimum of 1.6 mm.

CONCLUSIONSThe posterior and inferior quadrant of the footplate may be the optimal drill area for the fenestra.

Adult ; Humans ; Imaging, Three-Dimensional ; Saccule and Utricle ; anatomy & histology ; Stapes ; anatomy & histology ; Temporal Bone ; anatomy & histology

Objective To observe the protective effects of a mixture for protecting liver and supplementing stomach (保肝益胃合剂) on rat acute liver damage induced by carbon tetrachloride (CCl_4). Methods The model of rat acute liver damage was established by injection of CCl_4 2 ml/kg into the abdominal cavity.The rat models were treated respectively by the mixture for protecting liver and supplementing stomach 30 g?kg~(-1)?d~(-1),the polyene phosphatide acid radical choline capsule [Yi Shanfu (易善复), 180 mg?kg~(-1)?d~(-1)],the glycyrrhizic acid diaminogen capsule [Gan Lixin (甘利欣),30 mg?kg~(-1)?d~(-1)] infused into the stomach.The activities of serum alanine transaminase (ALT) and aspartate transaminase (AST) were detected.In the mean time,the liver pathological changes were observed,the degree of liver cell necrosis was evaluated,and the rat mortality was noted in various groups of treatment.Results The values of ALT,AST and the score of liver cell necrosis in the group treated with the mixture for protecting liver and supplementing stomach [(1.168?1.066) kU/L,(1.845?2.212) kU/L,(0.56?0.53) score] were significantly lower than those in the model group [(4.982?3.502) kU/L,(7.030?3.616) kU/L, (1.38?0.92) scores],and all the differences being statistically significant (all P

Objective To investigate the differences in the clinical symptoms and magnetic resonance imaging (MPd) findings of the spinal cord lesions between patients with neuromyelitis optica (NMO) and multiple sclerosis (MS) and explore the mechanisms that result in these differences. Methods The clinical symptoms and MRI findings of the spinal cord were retrospectively analyzed in 21 MS patients and 23 NMO patients admitted in the Third Affiliated Hospital of Sun Yat-Sen University from January, 2004 to January, 2007. Results Female patients were more frequently affected by NMO, and the NMO patients had a older mean age at onset with higher Expanded Disability Status Scale (EDSS) score than the MS patients. Chi-square test showed significant differences in the rotes of bilateral deep sensory dysfunction, zonesthesia, and sphincter disturbance between the NMO and MS patients (P<0.05). The majority of these clinical symptoms were found to result form corresponding spinal cord lesions defined by MRI. Conclusion NMO is a demyelinating disease that represents an independent clinical entity from MS, and has special mechanisms responsible for its characteristic clinical symptoms and MRI findings of the spinal cord.

OBJECTIVETo observe the effects of Tongxinluo Supermicro Powder on the nuclear factor-kappaB (NF-kappaB), inter-cell adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) expression in aorta of rabbits fed with high-lipid diet.

METHODSHealthy male New Zealand rabbits were randomly divided into 4 groups (n = 8 each): control group, model group, atorvastatin group (3 mg x kg(-1) x d(-1) per gavage), and Tongxinluo group (0.31 g x kg(-1) x d(-1) per gavage). At the end of 6 weeks, the expression of NF-kappaB, ICAM-1 and VCAM-1 were observed by immunochemistry methods, Western blotting and reverse transcription polymerase chain reaction (RT-PCR).

RESULTThe nuclear translocation of NF-kappaB in aortic endothelial cells and the gene expressions of NF-kappaB, ICAM-1 and VCAM-1 at protein and mRNA levels of the model group was significantly increased compared that in the control group (all P < 0.05), these effects could be significantly attenuated by atorvastatin and Tongxinluo Supermicro Powder (P < 0.01 vs. model group).

CONCLUSIONSSimilar as atorvastatin, Tongxinluo Supermicro Powder could relieve the process of atherosclerosis by decreasing the nuclear translocation of NF-kappaB and reducing the expression of ICAM-1, VCAM-1 in this model.

Animal Feed ; Animals ; Aorta ; drug effects ; metabolism ; Dietary Fats ; adverse effects ; Drugs, Chinese Herbal ; pharmacology ; Intercellular Adhesion Molecule-1 ; metabolism ; Male ; NF-kappa B ; metabolism ; Rabbits ; Vascular Cell Adhesion Molecule-1 ; metabolism

OBJECTIVETo determine the operation procedure of total hip arthroplasty (THA) for Crowe type-IV developmental dysplasia of hip and its relationship with nerve injury.

METHODSA consecutive series of 39 THAs was performed for Crowe type-IV developmental dysplasia of hip in 35 patients (all female). The mean age of the patients at the time of surgery was 46 years (range 36 - 56 years). Thirty-five hips in 31 patients were followed up. The average follow-up period of the whole series was 4 years (range 1 - 8 years). All procedures were carried out through a lateral-posterior approach. In 33 of 35 hips, the cup was inserted in the "true" acetabulum. All the prostheses used were cementless, except for 5 cemented femoral stems in 5 patients. Each patient was evaluated clinically and by radiographs before the operation and during the follow-up period, according to the Harris hip score (HHS).

RESULTSNone of the cups and stems were revised for aseptic loosening, dislocation or infection during the follow-up period. The mean preoperative HHS was 43 compared with the postoperative HHS of 87. The mean amount of postoperative leg lengthening was 5 cm (range 4 - 6 cm).

CONCLUSIONSThe reconstruction of the hip at the level of the "true" acetabulum through a lateral-posterior approach is a safe and effective procedure of THA for Crowe type-IV developmental dysplasia of hip in adults. Acute leg lengthening of less than 6 cm could not cause nerve injury.

Adult ; Arthroplasty, Replacement, Hip ; methods ; Female ; Follow-Up Studies ; Hip Dislocation, Congenital ; surgery ; Humans ; Middle Aged ; Treatment Outcome

BACKGROUNDThis study was designed to obtain a recombinant retroviral vector containing the human hepatocellular carcinoma-related gene ANGPTL4 (angiopoietin-like 4) cDNA and to evaluate the anti-tumor effect of recombinant retroviral vector-mediated human ANGPTL4 gene transfection.

METHODSANGPTL4 cDNA was cloned in vitro from normal human liver cells HL-7702 by using RT-PCR, and then subcloned into the plasmid vector pMSCV and sequenced. The retroviral plasmid vectors pMSCV-ANGPTL4, pVSV, and pGAG-POL were co-transfected into the packaging cell line 293 EBNA under mediation of lipofectamine. A high-titer retrovirus was obtained as a result, and HepG2 cells were infected with this retrovirus in vitro. Flow cytometry and fluorescence microscopy were used to detect expression of green fluorescence protein (GFP). The expression of ANGPTL4 mRNA in HepG2-ANGPTL4 cells was investigated using RT-PCR. The formation of tumors in nude mice and MTT assays were used to detect the growth of HepG2-ANGPTL4 cells in vivo and in vitro, respectively.

RESULTSThe cDNA sequence of the cloned ANGPTL4 gene was consistent with the recently reported sequence. Thus, the recombinant retroviral vector pMSCV-ANGPTL4 was constructed successfully. The titer of the packaged recombinant retrovirus was 1.4 x 10(6) infective viral grains/ml, and the rate of HepG2-ANGPTL4 cells expressing GFP was 68.45%, with an average intensity of fluorescence 31.67 times greater in HepG2-ANGPTL4 cells than in HepG2 cells. The expression of ANGPTL4 mRNA in HepG2-ANGPTL4 cells was higher than in HepG2-pMSCV cells (154% higher) or HepG2 cells (161% higher). The proliferation rate of HepG2-ANGPTL4 cells in vitro was obviously lower than those of HepG2-pMSCV cells and HepG2 cells (P <0.01). The mean volume and weight of tumors seeded from HepG2-ANGPTL4 cells were obviously lower than the mean volume or weight of tumors seeded from HepG2 cells and HepG2-pMSCV cells (P <0.01).

CONCLUSIONA stable ANGPTL4-transfected human liver cancer cell line HepG2-ANGPTL4 has been created. The transfer of the human ANGPTL4 gene mediated by a retroviral vector is a possibly effective approach for liver cancer therapy.

Angiopoietin-like 4 Protein ; Angiopoietins ; Animals ; Genetic Therapy ; Genetic Vectors ; genetics ; Humans ; Intercellular Signaling Peptides and Proteins ; genetics ; Liver Neoplasms ; therapy ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; NIH 3T3 Cells ; RNA, Messenger ; analysis ; Recombination, Genetic ; Retroviridae ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

OBJECTIVETo analyze related factors which affect GPA mutation frequency of workers exposed to benzene, with the Glycophorin A (GPA) mutation assay and explore the possibility of GPA mutation frequency as an index of predicting the risk of benzene poisoning.

METHODSThe erythrocytes were bound with fluorescent-labeled monoclonal antibody after isolated and fixed from the peripheral blood, and then the GPA mutation assay was performed using the flow cytometry (FCM). The related factors of GPA mutation frequency were analyzed by statistical methods.

RESULTSThe GPA mutation frequency of chronic benzene poisonings was significantly higher than that of their controls (P < 0.05). Significant direct correlation was found between age, length of service, accumulative exposure score and the GPA mutation frequency of workers exposed to benzene (P < 0.01). However, there was significantly inverse correlation between the 3AB index and the GPA mutation frequency (GPAN0: r(s) = -0.589, P < 0.01, GPANN: r(s) = -0.615, P < 0.01). In the multiple factor regression analysis on GPA mutation frequency, benzene exposure and individual susceptibility both entered model of multiple factors analysis, the coefficient of determination of benzene-exposed workers was 0.819.

CONCLUSIONExposure to benzene and individual susceptibility are the most important factors that affect GPA mutation frequency. GPA mutation frequency increases with the benzene exposure and individual susceptibility.

Benzene ; poisoning ; Glycophorin ; genetics ; Humans ; Mutation ; Mutation Rate ; Occupational Exposure