Objective To investigate whether miR-122 is a target of oxymatrine against HBV. Methods HepG2.2.15 cells were incubated with culture medium containing different concentrations of oxymatrine. The cyto-toxicity of oxymatrine was determined by cck-8 assay. The surface antigen of HBV (HBsAg),antigenof HBV (HBeAg)and HBV DNA in supernatant and intracellular miR-122 were determined in HepG2.2.15 cells after incu-bation with culture medium containing oxymatrine for 72 h. Results The survival percentage of HepG2.2.15 cells under different concentrations of oxymatrine was higher than 95% when the concentration of oxymatrine was lower than 4 mg/mL. After treatment with oxymatrine for 72 h,the secretion of HBsAg and HBeAg,the level of HBV DNA in the supernatant were reduced. The intracellular miR-122 in oxymatrine experience group was 3.5 times higher than that in the negative control group. Conclusions Oxymatrine might be used to inhibit viral replication and antigen expression by enhancing the expression of miR-122 in HepG2.2.15 cells.

OBJECTIVETo analyze the effects of three surgical operations in the treatment of Pilon fracture of Rüedi-Allgower type III, and put forward the best therapeutic method.

METHODSThe clinical data of 33 patients with Pilon fracture who received surgical operations (plaster immobilization group, 10 cases; distal tibia anatomical plate group, 11 cases; external fixation with limited internal fixation group, 12 cases) from October 2009 to January 2012 were analyzed. There were 5 males and 5 females, ranging in age from 24 to 61 years in the plaster immobilization group. There were 7 males and 4 females, ranging in age from 21 to 64 years in the distal tibia anatomical plate group. There were 7 males and 5 females, ranging in age from 23 to 67 years in the external fixation with limited internal fixation group. The Ankle X-ray of Pilon fracture after operation, ankle score, early and late complications were collected. Bourne system was used to evaluate ankle joint function.

RESULTSAfter 8 months to 3 years follow-up, it was found that three kinds of treatment had significant differences in the outcomes and complications (P < 0.05): the external fixation with limited internal fixation group got the best results. The number of anatomic reduction cases in the external fixation with limited internal fixation group (7 cases) and the distal tibia anatomical plate group (8 cases) was more than the plaster immobilization group (2 cases). According to the ankle score, 8 patients got an excellent result, 3 good and 1 poor in the limited internal fixation group ,which was better than those of distal tibia anatomical plate group (5 excellent, 4 good and 2 poor) and the plaster immobilization group (3 excellent, 4 good and 3 poor). The number of early and late complications in the external fixation with limited internal fixation group was more than those in the plaster immobilization group and the distal tibia anatomical plate group (P< 0.05).

CONCLUSIONTreatment of external fixation with limited internal fixation in the treatment of Pilon fracture of Rüedi-Allgower type III is effective and safe.

Adult ; Aged ; Case-Control Studies ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Tibial Fractures ; diagnostic imaging ; surgery ; Tomography, X-Ray Computed ; Treatment Outcome ; Young Adult

Objective To evaluate the clinical results of free vascularized fibular associated with iliac graft in treatment of old femoral neck fracture.Methods From 1994 to 1997,76 cases of old femoral neck fracture were treated with free vascularized fibular associated with iliac graft,they were followed up for observ- ing fracture healing,hip function and femoral head necrosis.Results Sixty-eight of these 76 cases were fol- lowed up with detail data with the main term as 10.5 years.All fractures healed within 4 to 6 months.63 cases got a almost normal hip function with the Harris score was 87.5,in these cases,the structure of femoral head were normal and the grafted fibular and iliac bone healed with the femoral.4 cases got to femoral head necrosis and 1 experienced hip dislocation.Conclusion Free vascularixed fibular associated with iliac graft is a good method to treat these cases with old femoral neck fracture who want reserve their femoral head.

OBJECTIVETo explore the effects of qianggu capsules (QGC) on the fracture healing and the bone mineral density (BMD) in radius distal osteoporosis fracture (RDOF) patients.

METHODSBone mineral density (BMD) of femoral neck in 65 patients with RDOF was detected after the fracture was fixed manually. They were then randomly divided into two groups. Thirty-three patients in the treated group took QC, 1 capsule (180 mg) each time, three times a day, while 32 patients in the control group took D-Cal Biocal 2 tablets (1500 mg) each time, once daily. The therapeutic course for both groups was three months. X-ray examination on the broken end of the fractured bone was taken every month to observe the bony callus formation for comparing the curative effect, and BMD of femoral neck were detected again after patients were treated for 3 months. The bony callus appeared earlier, more in volume with thicker cortex in the treated group after 2 months of treatment versus that in the control group. The fracture healing time in the treated group was 9.4 +/- 2.5 weeks and that in the control group was 12.5 +/- 2.9 weeks, showing significant difference between them (P < 0.05). BMD in the treated group before treatment was 0.621 +/- 0.085 g/cm2, which was lower than that after treatment (0.646 +/- 0.090 g/cm2) with significant difference showing between them (P < 0.05), while no significant change of BMD was found in the control group between before and after treatment, and significant difference was found in BMD between the two groups after treatment (P < 0.05).

CONCLUSIONQGC can promote the formation of bony callus ahead of time, increase the volume of bony callus and BMD, improve the bone structure, and thus the time of external fixation in treating RDOF could be reduced.

Aged ; Bone Density ; Capsules ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Fracture Healing ; drug effects ; Humans ; Male ; Middle Aged ; Osteoporosis ; complications ; Phytotherapy ; Radius Fractures ; complications ; drug therapy

OBJECTIVETo assess the effect of puerarin, a natural flavonoid found in Chinese Pueraria Lobata (Wild.) Ohwi, on promotion of new bone formation.

METHODSOsteoblasts isolated from calvarial of newborn rats were cultured in vitro in the presence of puerarin at various concentrations. The viability of osteoblasts and alkaline phosphotase activity and mineral node formation were determined. In addition, osteoblasts seeded in the β-tricaclium phosphate scalfolds as bone substitute were implanted in rat dorsal muscles. Half -of the recipient rats received intramuscular injection of puerarin at 10 mg/(kg·d) for 7 days. Osteogenesis was analyzed by examining the histology after 4 weeks of implantation.

RESULTSThe viability of osteoblasts treated with puerarin at either 40 or 80 μmol/L was significantly higher than that of the control (P<0.05 and P<0.01, respectively). Alkaline phosphatase and mineral modules were significantly increased in osteoblasts cultured with puerarin at 40 or 80 mol/L when compared with that of the untreated cells. The puerarin-treated rats had a higher rate of bone formation in the osteoblast implants than the control rats (6.35% vs. 1.32%, respectively, P<0.05).

CONCLUSIONPuerarin was able to affect osteoblast proliferation and differentiation, and promote the new bone formation in osteoblast implants.

Alkaline Phosphatase ; metabolism ; Animals ; Calcification, Physiologic ; drug effects ; Cell Differentiation ; drug effects ; Cell Survival ; drug effects ; Implants, Experimental ; Isoflavones ; pharmacology ; Male ; Microscopy, Electron, Scanning ; Osteoblasts ; cytology ; drug effects ; enzymology ; Osteogenesis ; drug effects ; Rats ; Rats, Sprague-Dawley ; Tissue Scaffolds

BACKGROUND: Osteoporosis, known as "a quiet epidemic disease", is an "invisible killer" that impairs the health of the elderly. Fracture as the most dangerous complication of osteoporosis is not only a medical problem, but also a serious social problem. OBJECTIVE: To summarize the selection, establishment methods, characters and evaluation of the animal model of osteoporosis. METHODS: A computed-based search of CBM, WanFang, VIP, CNKI and PubMed databases was conducted for the relevant articles published from January 1986 to December 2016 using the keywords of "osteoporosis, animal models" in English and Chinese, respectively. Finally 39 articles were included for result analysis after reading the title and abstract. RESULTS AND CONCLUSION: Animal experiments play an important role in the studies of osteoporosis, so a desirable animal model should hold similarity, reliability, repeatability, applicability, controllability, efficacy and economy. Osteoporosis is a progressive and long-lasting disease that has different characteristics at different stages. But an animal model only simulates one etiology, one stage, major symptoms and pathophysiological changes, and cannot exhibit the whole pathological changes. The current study suggests that animal species and modeling methods can affect the characters of models, and there is a lack of standards for animal age, feeding methods, modeling time and assessment.

Objective To investigate whether mitogen-activated protein kinases(MAPKs),which were involved in changes in osmolali-ty,might mediate aquaporin-4(AQP4)expression in astrocytes after oxygen-glucose deprivation(OGD)in rats.Methods Astrocytes were obtained from new born Sprague-Dawley rats.The P2cells were divided into control group,OGD group and inhibitors of U0126,SB203580 and SP600125 groups. The latter groups underwent OGD for five hours and reoxygenated, the inhibitors of U0126, SB203580 and SP600125(1 μmol/L and 10 μmol/L,respectively)groups,named U1,U10,SB1,SB10,SP1 and SP10 groups,respectively,were cultured with the inhibitors for twelve hours.The volume of cells in OGD group was measured half,one,one and half,two,three,four,eight and twelve hours after reoxygenation,and the expression of AQP4 was detected half,one,one and half,eight and twelve hours after reoxygen-ation with Western blotting.The expression of AQP4,and phosphorylation of extracellular regulated protein kinases(p-ERK),c-Jun N-termi-nal kinase(p-JNK)and p38 MAPK(p-p38 MAPK)was detected 24 hours after reoxygenation in all the groups,while the activity of lactate dehydrogenase(LDH)was measured.Results The volume of cells increased in OGD group one and half,two,three and four hours after re-oxygenation compared with those in the control group(P<0.001),and the expression of AQP4 also increased in OGD group after reoxygen-ation(P<0.001),especially 1.5 hours of reoxygenation.The expression of AQP4,p-ERK,p-JNK and p-p38 MAPK increased in OGD group after five hours of OGD compared with those in the control group(P<0.001),and the expression of p-ERK,p-JNK and p-p38 MAPK de-creased in the inhibitors groups compared with those in OGD group(P<0.001),and the expression of AQP4 decreased in SB10 group(P<0.001).The activity of LDH was less in all the inhibitors groups except SP1 and SB1 groups than in OGD group(P<0.01),and was the least in SB10 group (P<0.001). Conclusion MAPKs signal pathway, especially p38 MAPK, may promote AQP4 expression in astrocytes after OGD in rat,and play a role in cells death.

OBJECTIVETo express human vascular endothelial growth factor (hVEGF(165)) in E. coli JM109 in the form of fusion protein by genetic engineering and test the biological activity and immunological competence of the expressed protein.

METHODShVEGF(165) gene was subcloned by PCR and inserted into pQE30 plasmid. hVEGF(165) fusion protein was expressed in E. coli JM109 and purified by Ni(2+)-NTA. The immunological competence of the expressed protein was tested by means of Western blotting and enzyme-linked immunosorbent assay (ELISA), and its biological activity was assayed by chicken chorioallantoic membrane (CAM) and Matrigel angiogenesis assay.

RESULTSThe recombinant hVEGF(165) fusion protein was successfully expressed in E. coli JM109 and its expression accounted for 30% of the total cellular protein. The purified protein presented a single band of 23 kD in SDS-PAGE. Western blotting, ELISA, CAM and matrigel angiogenesis assay showed excellent immunologic competence and biological activity of the recombinant protein.

CONCLUSIONRecombinant hVEGF(165) protein with excellent biological activity has been successfully expressed in E.coli JM109, which may facilitate future study in construction of prefabricated tissue-engineered bone graft.

Blotting, Western ; Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; genetics ; metabolism ; Humans ; Plasmids ; genetics ; Prokaryotic Cells ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification ; Vascular Endothelial Growth Factor A ; biosynthesis ; genetics ; isolation & purification

OBJECTIVETo evaluate stress changes of intervertebral space and adjacent intervertebral space after artificial disc replacement with angles.

METHODSArtificial disc replacement with angles were designed according to existing data. Axial pressure, flexion/extension, lateral bending and torsion loading were applied on finite element models of normal cervical discs on C4,5 segments, C4,5 segments with 0 degrees artificial cervical discs and C4,5 segments with 10 degrees artificial cervical discs, then stress changes of C4,5 space was observed. The same loadings were applied on finite element models of normal cervical discs on C4-C6 segments, C4,5 segments with 0 degrees, C4,5 segments with 10 degrees, then stress changes of replaced segments space and adjacent segment space were observed.

RESULTSFor C4,5 segments, 80 MPa/0 degrees artificial discs and 80 MPa/10 degrees artificial discs had the similar equivalent shear stress (Se), and were both larger than that of normal discs, when lateral bending were performed, 80 MPa/0 degrees artificial discs were closed to normal discs when axial pressure and flexion/extension were carried out, while 80 MPa/10 degrees artificial discs had a larger Se than that of normal ones,when torsion loading were applied, Szx/Szy stress of 80 MPa/0 degrees and 80 MPa/10 degrees artificial discs were closed to normal ones. For C4-C6 segments, the axial pressure, flexion/extension and lateral bending of C5,6 were all lower than normal discs after C4,5 discs were replaced by 80 MPa/10 degrees artificial discs, while Szx/Szy of torsion loading were closed to normal ones.

CONCLUSIONArtificial discs with 10 degrees have less influences on stress of adjacent intervertebral space and closer to mechanical property after being implanted into intervertebral space.

Cervical Vertebrae ; surgery ; Humans ; Stress, Mechanical ; Total Disc Replacement ; methods

BACKGROUNDChinese medicine plays an important role in hepatoprotective treatment. This study was conducted to investigate the protective effects of emodin and astragalus polysaccharides (APS) in a rat model of chronic hepatic injury.

METHODSChronic hepatic injury was induced by hypodermic injection of an olive oil solution containing 40% carbon tetrachloride (CCl(4)) twice a week, in addition to a diet of 79.5% maizena, 20% fat, 0.5% cholesterol, and 10% alcohol in the drinking water ad libitum for 12 weeks. Meanwhile, the rats were exposed to different concentrations of emodin (40 mg x kg(-1) x d(-1)), APS (200 mg x kg(-1) x d(-1)), combination drug (emodin 40 mg x kg(-1) x d(-1) combined with APS 200 mg x kg(-1) x d(-1)) and colchicine (0.1 mg x kg(-1) x d(-1)) in parallel by oral gavage (once a day for 12 weeks). At the end of 12 weeks, blood serum and liver tissue were taken. Serum was collected to determine the levels of total bilirubin (TBIL), alanine transaminase (ALT), aspartate transaminose (AST), and albumin (ALB). Liver and spleen indexes were assayed, followed by the measurements of the liver associated enzyme superoxide dismutase (SOD) and malondialdehyde (MDA). Histopathological changes were studied using optical microscopy.

RESULTSSplenohepatomegalia was alleviated and serum levels of TBIL and ALT were reduced in the groups treated with emodin and APS when compared to the control group. In addition, the ALB level in the APS and combination groups was higher. Similarly, the SOD activity of liver homogenates was significantly higher in the groups treated with emodin and APS, while administration of the herbal derivatives prevented the elevation in MDA levels. Histological analysis showed that the APS and combination groups significantly ameliorated the hepatic injury.

CONCLUSIONSCo-administration of emodin and APS demonstrated a synergistic action in reducing ALT and restoring ALB in the serum from a rat model of chronic hepatic injury. Emodin and APS may ameliorate the CCl(4)-induced hepatic injury in rats by elevating antioxidant-enzyme activities and reducing lipid peroxidation.

Alanine Transaminase ; blood ; Animals ; Astragalus Plant ; chemistry ; Carbon Tetrachloride ; toxicity ; Chronic Disease ; Emodin ; pharmacology ; Liver ; drug effects ; pathology ; Male ; Malondialdehyde ; analysis ; Polysaccharides ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism