Objective To investigate the effect of Sclareol(SCL)on LX-2 hepatic stellate cell activation and CCl4-induced liver fibrosis in mice,and to further explore its mechanism.Methods A total of 40 Kunming mice were randomly divided into healthy group,model group(10%CCl4)and SCL administration group,and silybin positive control group(10%CCl4+100 mg·kg-1 Silybin),and SCL administration group was divided into low SCL(10%CCl4+20 mg·kg-1 SCL)and high dose group(10%CCl4+40 mg·kg-1 SCL).Mice in all groups were intraperitoneally injected with 10%olive oil-diluted CCl4 three times a week for four weeks,except for the healthy group.Starting from the third week,the dosing group was given different doses of SCL by gavage daily,and the positive control group was given silybin daily,and the mice were sacrificed and serum and liver tissue were collected after four weeks.In whole animal experiments,biochemical kits were used to detect the changes in the serum levels of glutamate aminotransferase(ALT)and aspartate aminotransferase(AST)in mice with liver fibrosis.Hematoxylin-eosin(HE),Sirius Red and Masson staining were used to detect microstructural changes and collagen deposition in liver tissues.Immunohistochemistry was used to detect the expression of fibrosis marker proteins α-smooth muscle actin(α-SMA)and fibrous collagen I.in liver tissues.In vitro,LX-2 human hepatic stellate cells were used for normal culture in the blank group,and the activation of LX-2 hepatic stellate cells was induced by transforming growth factor-β1(TGF-β1)in the model group,and the SCL administration group was divided into SCL low-dose group(5 ng·mL-1 TGF-β1+10 μmol·L-1 SCL)and high-dose group(5 ng·mL-1 TGF-β1+20 μmol·L-1 SCL).Subsequently,Transwell and EdU assays were used to detect the effects of SCL on the migration and proliferation of LX-2 cells.The expression of fibrosis marker proteins α-SMA and Collagen I.affected by SCL was detected by immunofluorescence.Western blot was used to detect the expression of related proteins in TGF-β/Smad pathway.Results In animal experiments,compared with the model group,SCL could significantly improve the liver function indexes and liver histopathological changes in liver fibrosis model mice.In addition,in vitro cell experiments,compared with the model group,SCL can effectively inhibit the migration and proliferation of hepatic stellate cells and inhibit their activation.Further studies showed that compared with the model group,SCL significantly up-regulated the expression of Smad7 protein and significantly down-regulated the phosphorylation levels of Smad2 and Smad3 proteins.Conclusion SCL has a significant alleviating effect on CCl4-induced liver fibrosis and TGF-β1-induced LX-2 activation in mice,and the mechanism may be related to the regulation of TGF-β/Smad pathway.

Objective To investigate the effect of Sclareol(SCL)on LX-2 hepatic stellate cell activation and CCl4-induced liver fibrosis in mice,and to further explore its mechanism.Methods A total of 40 Kunming mice were randomly divided into healthy group,model group(10%CCl4)and SCL administration group,and silybin positive control group(10%CCl4+100 mg·kg-1 Silybin),and SCL administration group was divided into low SCL(10%CCl4+20 mg·kg-1 SCL)and high dose group(10%CCl4+40 mg·kg-1 SCL).Mice in all groups were intraperitoneally injected with 10%olive oil-diluted CCl4 three times a week for four weeks,except for the healthy group.Starting from the third week,the dosing group was given different doses of SCL by gavage daily,and the positive control group was given silybin daily,and the mice were sacrificed and serum and liver tissue were collected after four weeks.In whole animal experiments,biochemical kits were used to detect the changes in the serum levels of glutamate aminotransferase(ALT)and aspartate aminotransferase(AST)in mice with liver fibrosis.Hematoxylin-eosin(HE),Sirius Red and Masson staining were used to detect microstructural changes and collagen deposition in liver tissues.Immunohistochemistry was used to detect the expression of fibrosis marker proteins α-smooth muscle actin(α-SMA)and fibrous collagen I.in liver tissues.In vitro,LX-2 human hepatic stellate cells were used for normal culture in the blank group,and the activation of LX-2 hepatic stellate cells was induced by transforming growth factor-β1(TGF-β1)in the model group,and the SCL administration group was divided into SCL low-dose group(5 ng·mL-1 TGF-β1+10 μmol·L-1 SCL)and high-dose group(5 ng·mL-1 TGF-β1+20 μmol·L-1 SCL).Subsequently,Transwell and EdU assays were used to detect the effects of SCL on the migration and proliferation of LX-2 cells.The expression of fibrosis marker proteins α-SMA and Collagen I.affected by SCL was detected by immunofluorescence.Western blot was used to detect the expression of related proteins in TGF-β/Smad pathway.Results In animal experiments,compared with the model group,SCL could significantly improve the liver function indexes and liver histopathological changes in liver fibrosis model mice.In addition,in vitro cell experiments,compared with the model group,SCL can effectively inhibit the migration and proliferation of hepatic stellate cells and inhibit their activation.Further studies showed that compared with the model group,SCL significantly up-regulated the expression of Smad7 protein and significantly down-regulated the phosphorylation levels of Smad2 and Smad3 proteins.Conclusion SCL has a significant alleviating effect on CCl4-induced liver fibrosis and TGF-β1-induced LX-2 activation in mice,and the mechanism may be related to the regulation of TGF-β/Smad pathway.

OBJECTIVE To explore the protective effect of polysaccharides from Salvia Chinensis Benth. (PSSC) on lipopolysaccharide (LPS) and D- galactosamine (GalN)- provoked mouse acute liver failure (ALF) and the possible molecular mechanism. METHODS Kunming mice were randomly divided into four groups: normal control, model, model+PSSC 30 and 100 mg·kg- 1 groups. PSCC was given once a day and for a week. To establish an ALF model, mice of model and PSSC groups were ip injected with LPS 10 μg·kg-1 and GalN 700 mg·kg-1 at the end of PSSC treatment. The microscopic structure of the liver was detected by HE staining. Serum and hepatic biochemical parameters of glutamic-oxalacetic transaminase (GOT), glutamic- pyruvic transaminase (GPT), catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), malondialdehyde (MDA), and glutathione (GSH) were detected by colorimetric methods. The relative content of hepatic reactive oxygen species (ROS) was measured by DCFH-DA fluorescent probes. Levels of cytokines tumor necrosis factor-α (TNF-α), interleukin 1β(IL-1β) and IL- 6 in the serum and liver were detected by ELISA. Activity of caspase 3 in liver homogenates was detected by aspase 3 activity assay kit. RESULTS Compared with normal control group, in the liver of model group, hepatocytes were arrayed in disorder, cytoplasm of hepatocytes shrank, and boundaries between cells were fuzzy, the infiltration of a large number of inflammatory cells and tissue hemorrhage could be detected, pathological scores were elevated significantly (P<0.01), levels of MDA and ROS in the liver of ALF model mice were elevated to 2.2 and 4.3 times that of the normal control, respectively (P<0.01), the level of GSH decreased to 51% (P<0.01), and the activities of SOD, CAT and GSH- Px declined to 74%, 36% and 42%, respectively (P<0.01). Levels of TNF- α, IL- 1β and IL- 6 in the serum and liver of model group were increased (P<0.01), and caspase 3 activity was increased to 5.3 times that of the normal control (P<0.01). Compared with the model group, the number of surviving mice in PSSC groups increased, liver pathological scores declined (P<0.01), levels of MDA and ROS increased (P<0.01), levels of GSH, TNF-α, IL-1β and IL-6 in the liver and serum declined (P<0.01), and caspase 3 activity decreased (P<0.01). CONCLUSION PSSC is able to alleviate LPS and GalN-induced ALF in mice. Inhibition of hepatic oxidative stress, inflammatory response, and cell apoptosis is possibly implicated in the protective effect of PSSC.

Objective To evaluate the clinical outcomes and correction accuracy of posttraumatic varus and valgus knee deformities using the Taylor Spatial Frame (TSF).Methods From June 2006 to December 2010,26 patients with posttraumatic varus and valgus knee deformities who had been treated with the TSF combined with less invasive high tibial osteotomy (HTO) or supracondylar femur osteotomy (SFO) were retrospectively analyzed,including 19 males and 7 females with an average age of 39 years.There were 20 cases of posttraumatic varus knees and 6 valgus knees.Twenty one patients were treated with HTO and 3 with SFO,2 with HTO and SFO simultaneously.During the operation,the TSF was firstly assembled and then the parameters of the frame were measured followed by less invasive percutaneous osteotomy.Correction was started seven to ten days after the operation following the time schedule provided by the computer program.Correction of the struts were performed 3 times per day,with a total movement of 0-3 mm.Results Through 7-35 days frame adjustments,20 cases of deformities were fully corrected.X-ray showed that the mechanical axis deviations of the tibia and femur all were recovered to normal range,and the deformity of rotation,angulation and crispation were fully corrected.Six cases had residual slight angulation or crispation deformity ( ＜ 4° or ＜ 10 mm).Through the second 4-10 days frame adjustments,these 6 cases of deformities were fully corrected finally.New bone formation and consolidation on the osteotomy site were good.The fixators were removed from 2.5-6.0 months after surgery and the appearance of the limbs and the patients' gait recovered to normal and the keen pain disappeared.All patients were followed up for 12-60 months and without the recurrence of the deformity.Eleven cases occured pin site infection and were controlled by use of the sensitive antibiotics.One case occured refracture of the distal femur osteotomy site 1.5 months after frame removal.Through conservative treatment,the fracture finally healed.Conclusion Posttraumatic varus and valgus knee deformities can be effectively corrected by using TSF and with a high accuracy.

OBJECTIVETo study the effect of total saponins from Entada phaseoloides (TSEP) on islet morphology and skeletal muscle PI3K pathway-related protein expression of type 2 diabetic rats.

METHODType 2 diabetic rats were induced by high-fat diet and low-dose streptozotocin and then randomly divided into 5 groups, i.e. the normal control, the model group, the positive control drug (200 mg x kg(-1) metformin), the low-dose TSEP (25 mg x kg(-1)) group and the high-dose TSEP (50 mg x kg(-1)). Three weeks later, the islet morphology of rat pancreas were observed by HE staining, and protein expressions of insulin receptor substrate-1 (IRS-1), phosphatidylinositol 3-kinase (PI3K), protein tyrosine phosphatase-1B (PTP-1 B) and glucose transporter 4 (GLUT4) in rat skeletal muscle were detected by Western blot.

RESULTCompared with the modal group, TSEP administration groups showed relatively normal structures, clear pancreatic cells and intact capsula structures in pancreatic tissue pathological sections, with the number of pancreatic islets close to the normal control group. Meanwhile, above TSEP administration groups showed increased IRS-1, PI3K and GLUT4 protein expressions in their skeletal muscle tissues and decreased PTP-1B protein expression compared with the model group.

CONCLUSIONTSEP has an effect on protecting pancreatic tissues of type 2 diabetic rats and intervening in abnormal expression of proteins in skeletal muscle tissues.

Animals ; Diabetes Mellitus, Type 2 ; drug therapy ; Fabaceae ; chemistry ; Glucose Transporter Type 4 ; analysis ; Islets of Langerhans ; drug effects ; pathology ; Male ; Muscle, Skeletal ; drug effects ; pathology ; Phosphatidylinositol 3-Kinases ; analysis ; Rats ; Rats, Sprague-Dawley ; Saponins ; therapeutic use

OBJECTIVETo study in vitro anti-tumor activity of phaseoloideside E (PE) with human hepatoma HepG2 cells as the objective.

METHODMTT assay was adopted to detect the cytotoxic effect of PE of different concentrations on HepG2 cells after being processed for 48 h. Changes in morphology of PE-processed cells were observed under an optical microscope and fluorescence microscope. DNA agrose gel electrophoresis was used to detect the DNA ladder, an important characteristic of cell apoptosis. The expression levels of Bax and Bcl-2 were determined by western blot assay.

RESULTPE dramatically repressed the viability of HepG2 cells. Typical morphological changes of apoptosis had been detected by both direct microscopic observation and Hoechst 33,258 staining. Typical DNA Ladder was also observed by agarose gel electrophoresis in the administration group, but it did not exist in the control group. Western blot showed that the expression of Bax was up-regulated and Bcl-2 was down-regulated.

CONCLUSIONAbove data demonstrates that PE can induce apoptosis in human hepatoma HepG2 cells, and indicate that PE-induced expression level changes of Bax and Bcl2 may be related to the apoptosis-induction effect.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Hep G2 Cells ; Humans ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; Saponins ; pharmacology ; Triterpenes ; pharmacology ; bcl-2-Associated X Protein ; analysis

[Objective]To investigate the clinic application of the reversed sural neurocutaneous island flap in repairing the soft tissue defect of the foot and ankle.[Method]From June 1997 to December 2007,38 cases of the lower leg,around the ankle and the heel soft tissue defects were treated with the reversed sural neurocutaneous island flap.The average of the patients was 38.3 years(range 5 to 65).The longest medical history was 11 years.The lagest area of flap was 17 cm?12 cm and the smallest was 5 cm?4 cm.[Result]Thirty-three flaps were completely survived,there were 5 cases with distal edge of the flaps partially necrosis and healed after short time dressing change.The patients were followed up for half a year to 10 years and the curative effect was satisfactory.[Conclusion]The reversed sural neurocutaneous flap is simple to harvest and its blood supply is reliable without the sacrifice of a major blood vessel.So it is an ideal method for repairing the soft tissue defect of the foot and ankle.

[Objective]To observe the clinical effect of proximal humeral fractures treated with locking proximal humerus plate.[Method]From April 2004 to August 2008,21 patients with proximal humeral fractures were all treated with locking proximal humerus plates,the average age of the patients was 51 years(range 35-78).According to Neer classification:two-part fracture 7,three-part fracture 12,four-part fracture 2.[Result]Twenty-one patients were all followed up for 6-24 months(average 14 months),all fractures occurred bony union.Clinical effect was evaluated by Neer functional assessment system:excellent 14,good 5,fare 2.[Conclusion]Application of locking proximal humerus plates in treating proximal humeral fractures can obtain satisfactory therapeutic result and is a more ideal method of internal fixation of proximal humeral fractures at present,attaching importance to injury of shoulder sleeve and early exercise,most patients can acquire satisfactory clinical effect.