OBJECTIVE　To develop a method for the determination of theophylline in serum with concomitant use of ofloxacin.METHODS　Theophylline was serum was extracted with chloroform-hexane(7∶3,v/v) after some ammonium sulfate was added.It was re-extracted with NaOH solution (0.1 mol*L-1)and detected with a UV spectrophotometer at 275 nm and 299.5 nm respeitively.RESULTS　The method was linear over the range of 5.0～40.0 mg*L-1(r=0.9997,n=7).The average recovery of methodology was 100.3%(RSD=1.2%),and the extraction recovery from serum was 87.7%(RSD=4.6%).CONCLUSION　This practical method for the determination of serum theophylline can eliminate the interference of ofloxacin and other antibiotics.

Objective To investigate the effects of external fixator with dynamic device under low frequency and controlled micromovement on the callus calcification and mechanical property of healing bone.MethodsForty-five sheep were performed transverse osteotomy with a gap of 2 mm on the mid-shafts of both tibias,and the hind limbs were fixed with unilateral external fixators connected to a controlled micromovement device.Ten days after osteotomy,one hind limb of each sheep was randomly selected for micromovement(30 min/d).According to different micromovement frequencies,the sheep were randomly divided into 3 groups: 0.5 Hz group,1 Hz group and 5 Hz group(n=15).The amplitude of micromovement was 0.25 mm and the micromovement stopped by the end of the fourth week postoperation.The other hind limb of each sheep was served as control group without micromovement.Morphometry of callus was done at the end of 4,6 and 9 weeks after osteotomy.Bone formation velocity,bone mineral density and biomechanical properties were compared at the end of 9 weeks.Results The areas of mineralized bone and osteoid in different miromovement groups were larger than that of control group at the end of 4,6 weeks postoperation(P

To study the effect of the combined administration of different doses of Glycyrrhizae Radix et Rhizoma and Atractylodis Macrocephalae Rhizoma on the proliferation of DFMO-treated intestinal epithelial cells (IEC-6) and p53, p21 mRNA and protein expressions, in order to define the molecular basis for the effect of the combined administration of different doses of Glycyrrhizae Radix et Rhizoma and Atractylodis Macrocephalae Rhizoma on the cell proliferation. The effect of the drugs on the cell division rate and cell cycle of IEC-6 cells was detected by FCM. Quantitative Real-time PCR (qRT-PCR) was used to analyze the effect of the drugs on mRNA of p2l and p53 related to IEC-6 proliferation. Western blot was used to analyze the effect of the drugs on p2l and p53 protein expressions of IEC-6 cells. Atractylodis Macrocephalae Rhizoma could increase p53, p21 mRNA and proteins expression in DFMO-treated IEC-6 cells. The combined administration of different ratios of Atractylodis Macrocephalae Rhizoma and Glycyrrhizae Radix et Rhizoma could significantly down-regulate Atractylodis Macrocephalae Rhizoma's effect on p53, p21 mRNA and proteins expression in DFMO-treated IEC-6 cells and promote the proliferation of IEC-6 cells. The combined administration of Atractylodis Macrocephalae Rhizoma and Glycyrrhizae Radix et Rhizoma could down-regulate Atractylodis Macrocephalae Rhizoma's effect on DFMO-treated intestinal epithelial cells (IEC-6).
Animals ; Atractylodes ; chemistry ; Cell Line ; Cyclin-Dependent Kinase Inhibitor p21 ; genetics ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Epithelial Cells ; drug effects ; metabolism ; Gene Expression ; drug effects ; Glycyrrhiza ; chemistry ; Intestines ; drug effects ; metabolism ; Rats ; Rhizome ; chemistry ; Tumor Suppressor Protein p53 ; genetics ; metabolism

Animals ; Disease Models, Animal ; Liver ; parasitology ; Mice ; Mice, Inbred BALB C ; Schistosomiasis japonica ; immunology ; Thiourea ; analogs & derivatives ; pharmacology

OBJECTIVETo determine the safety and efficacy of local administration of lentivirus-mediated small interfering RNA (siRNA) targeting tumor necrosis factor-alpha (TNF-alpha) in murine air pouch model.

METHODSFrom May 2007 to April 2008 a siRNA targeting TNF-alpha and a missense siRNA were designed, and recombine lentivirus which coexpressed the green fluorescent protein (GFP) as a marker gene was constructed. Air pouches were established and stimulated by Ti-6Al-4V particles. Pouches were divided into 3 groups randomly. Lentivirus-mediated siRNA targeting TNF-alpha (TNF-alpha group) or lentivirus-mediated missense siRNA (MS group), or virus-free saline (control group) were injected into pouches respectively. Pouch membrane, peripheral blood, heart, liver, spleen, kidney, lung and brain were harvested at 28 d after transfection, and assayed for markers of inflammation using histological, molecular, immunological techniques and Xenogen in vivo imaging system (IVIS) 50 vivo bioluminescent assay system.

RESULTSXenogen IVIS 50 vivo image revealed strong expression of GFP localized in pouch areas and no expression in other parts of mice both in TNF-alpha group and MS group at 4 weeks after transfection, while no expression of GFP was found in control group. By RT-PCR and ELISA, the mRNA and protein levels of TNF-alpha in TNF-alpha group decreased by 81.6% and 82.6% respectively compared to control group (P < 0.01), and decreased by 78.9% and 84.0% respectively compared to MS group (P < 0.01), whereas TNF-alpha level in peripheral blood, heart, liver, spleen, kidney, lung and brain remained invariant (P > 0.05). Less inflammatory responses (thinner pouch membrane and decreased cellular infiltration) were observed in TNF-alpha group.

CONCLUSIONEfficient local delivery of lentivirus-mediated siRNA targeting TNF-alpha into modified murine air pouch can inhibit debris-induced inflammation effectively, with no systemic adverse effects.

Animals ; Disease Models, Animal ; Genetic Therapy ; Genetic Vectors ; genetics ; Inflammation ; therapy ; Lentivirus ; genetics ; Mice ; Mice, Inbred BALB C ; RNA, Small Interfering ; genetics ; Random Allocation ; Transfection ; Tumor Necrosis Factor-alpha ; genetics

BACKGROUNDTotal hip arthroplasty (THA) is widely applied for the treatment of end-stage painful hip arthrosis. Traditional THA needed a long incision and caused significant soft tissue trauma. Patients usually required long recovery time after the operation. In this research we aimed to study the feasibility and clinical outcomes of minimally invasive two-incision THA.

METHODSFrom February 2004 to March 2005, 27 patients, 12 males and 15 females with a mean age of 71 years (55 - 76), underwent minimally invasive two-incision THA in our department. The patients included 9 cases of osteoarthritis, 10 cases of osteonecrosis, and 8 cases of femoral neck fracture. The operations were done with VerSys cementless prosthesis and minimally invasive instruments from Zimmer China. Operation time, blood loss, length of incision, postoperative hospital stay, and complications were observed.

RESULTSThe mean operation time was 90 minutes (80 - 170 min). The mean blood loss was 260 ml (170 - 450 ml) and blood transfusion was carried out in 4 cases of femoral neck fracture (average 400 ml). The average length of the anterior incision was 5.0 cm (4.6 - 6.5 cm) and of the posterior incision 3.7 cm (3.0 - 4.2 cm). All of the patients were ambulant the day after surgery. Nineteen patients were discharged 5 days post-operatively and 8 patients 7 days post-operatively. The patients were followed for 18 months (13 - 25 months). One patient was complicated by a proximal femoral fracture intraoperatively. No other complications, including infections, dislocations, and vascular injuries, occurred. The mean Harris score was 94.5 (92 - 96).

CONCLUSIONSTwo-incision THA has the advantage of being muscle sparing and minimally invasive with less blood loss and rapid recovery. However, this technique is time consuming, technically demanding, and requires fluoroscopy.

Aged ; Arthroplasty, Replacement, Hip ; methods ; Female ; Fluoroscopy ; Humans ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; methods ; Retrospective Studies

OBJECTIVETo investigate the effect of PD98059 on the proliferation and cell cycle of rat hepatic stellate cells (HSCs) stimulated by acetaldehyde and explore its mechanism.

METHODSRat HSCs stimulated by acetaldehyde were incubated with different concentrations of PD98059. Cell proliferation was assessed by MTT colorimetric assay. Cell cycle was analysed by flow cytometry. The mRNA of cyclin D1 and CDK4 were examined by RT-PCR.

RESULTS20, 50, 100 micromol/L PD98059 could significantly inhibit the proliferation of HSCs stimulated by acetaldehyde in a does-dependent manner (0.109+/-0.020, 0.081+/-0.010 and 0.056+/-0.020 vs 0.146+/-0.030, F=31.385, P<0.05) and provoke G0/G1 phase arrest of HSCs stimulated by acetaldehyde in a does-dependent manner (61.9%+/-6.3%, 64.1%+/-3.3% and 70.9%+/-4.8% vs 55.2%+/-4.4%, F=16.402, P<0.05). 50, 100 micromol/L PD98059 could markedly inhibit cyclin D1 mRNA expression of HSC stimulated by acetaldehyde (0.56+/-0.04 and 0.46+/-0.03 vs 0.65+/-0.07, F=68.758, P<0.05) and CDK4 mRNA expression (0.39+/-0.07 and 0.33+/-0.05 vs 0.50+/-0.06, F=29.406, P<0.05).

CONCLUSIONThe Erk signal transduction pathway plays an important role in regulating the proliferation and cell cycle of rat hepatic stellate cells stimulated by acetaldehyde, which may be partly related to its regulative effect on the expression of cyclin D1 gene and CDK4 gene

Acetaldehyde ; pharmacology ; Animals ; Cells, Cultured ; Cyclin D1 ; metabolism ; Cyclin-Dependent Kinase 4 ; Cyclin-Dependent Kinases ; metabolism ; Enzyme Inhibitors ; pharmacology ; Flavonoids ; pharmacology ; Hepatocytes ; drug effects ; Proto-Oncogene Proteins ; Rats

OBJECTIVETo report the clinical outcome of minimally invasive total hip arthroplasty with anterior incision.

METHODSOne hundred and twenty cases were randomly divided into two groups. Sixty cases (group 1) who had undergone a mini-invasive THA were compared with 60 cases (group 2) who had undergone THA with standard posterolateral incision. The operation time, length of incision, blood loss, anteversion angle of acetabulum cup, Harris score and complications were observed.

RESULTThe average operation time was almost the same; The average length of incision for group 1 was 7.9 cm (7.4 - 9.0 cm) and 16.3 cm (14 - 22 cm) for group 2 (P < 0.01). The average blood loss for group 1 was 350 ml (250 - 530 ml) and 650 ml (400 - 1200 ml) for group 2, there was significant difference between two groups (P < 0.05). According to the postoperative X-ray, the mean anteversion angles of cup were 24 degrees (19 degrees - 27 degrees) for group 1 and 19 degrees (15 degrees - 22 degrees) for group 2. The average length of post-operative hospital stay was 7 days (5 - 8 days) in group 1 and 13.5 days (12 - 16 days) in group 2 (P < 0.05). The Harris score of group 1 was 91.4 (67 - 94) and 78.5 (67 - 91) for group 2 at the 3 month follow-up (P < 0.05). At the last follow up, there were no significant difference (P > 0.05), but the average ROM of group 1 were definitely more greater than that of group 2 (110.0 degrees +/- 3.2 degrees vs. 90.0 degrees +/- 2.9 degrees P < 0.05). There was 1 case of cup reinsertion in group 1 because of large anteversion angle; 2 cases of symptomatic DVT and 1 case of lethargy because of cerebral infarction happened in group 2.

CONCLUSIONSMinimally invasive total hip arthroplasty using anterior approach is a safe and effective technique with the advantages of less soft tissue damage and less blood loss.

Aged ; Arthroplasty, Replacement, Hip ; methods ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures

AIMTo study the effect of aspirin on chronic hypoxia and hypercapnic pulmonary hypertension.

METHODSSD rats were randomly divided into normal control group (A), hypoxic hypercapnic group (B), hypoxic hypercapnia + aspirin group (C). The concentration of TXB2 and 6-keto-PGF1alpha in plasma and in lung were detected by the technique of radioimmunology.

RESULTS(1) mPAP was significantly higher in B group than those of A and C group. Differences of mCAP were not significant in three groups. (2) Light microscopy showed that WA/TA (vessel wall area/total area) and PAMT (the thickness of medial smooth cell layer) were significantly higher in B group than those of A and C group. (3) The concentration of TXB2 and 6-keto-PGF1alpha in plasma and lung as well as the ratio of TXB2/6-keto-PGF1alpha were significantly higher in rats of B group than those of A and C group.

CONCLUSIONAspirin may inhibit hypoxic hypercapnia pulmonary hypertension and pulmonary vessel remodeling.

6-Ketoprostaglandin F1 alpha ; metabolism ; Animals ; Aspirin ; pharmacology ; Carotid Arteries ; pathology ; physiopathology ; Epoprostenol ; metabolism ; Hypercapnia ; physiopathology ; Hypertension, Pulmonary ; metabolism ; pathology ; physiopathology ; Hypoxia ; physiopathology ; Male ; Pulmonary Artery ; pathology ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Thromboxane A2 ; metabolism

AIMTo study the effect of chronic hypoxic hypercapnia on expression of COX-2 mRNA in pulmonary arterioles.

METHODSSD rats were randomly divided into two groups: control group and hypoxic hypercapnic group. COX-2 mRNA was observed in pulmonary arterioles by the technique of in situ hybridization.

RESULTSmPAP, weight ratio of right ventricle (RV) to left ventricle plus septum (LV + S) and COX-2 mRNA in pulmonary arterioles were much higher in rats of hypoxic hypercapnic group than those of control group. Light microscopy showed that vessel smooth muscle cell hypertrophy and vessel cavity straightness were found in hypoxic hypercapnic group.

CONCLUSIONChanges of expressions of COX-2 mRNA may regulate hypoxic hypercapnic pulmonary hypertension.

Animals ; Cyclooxygenase 2 ; genetics ; metabolism ; Hypercapnia ; metabolism ; physiopathology ; Hypoxia ; metabolism ; physiopathology ; Male ; Pulmonary Artery ; metabolism ; physiopathology ; Rats ; Rats, Sprague-Dawley