Objective To purify ?-glutamyltranspeptidase (GGT) strongly combined to datura stramonim (DSA) lectin from human liver cancer tissue,which will be used as the antigen to produce antibodies for early diagnosis of liver cancer.Methods The GGT strongly combined to DSA lectin in human liver cancer tissue was purified by lectin affinity chromatography from human liver cancer tissue, including rough extraction,dialysis,ion-exchange chromatography and lectin affinity chromatography.Results The catalysis units per milligram protein of GGT increased about 84.9 times.The yield of the GGT was 1.60%.Conclusion The protocol in this study is efficient to purity GGT from human liver cancer tissue.

BACKGROUND:Silk fibroin, as an inorganic mineralization template, can induce hydroxyapatite crystal growth, and combined with nano-hydroxyapatite can simulate the inorganic and organic components of natural bone, which is expected to become the most ideal bone graft material. OBJECTIVE:To prepare the silk fibroin/nano-hydroxyapatite composite material and investigate its treatment outcomes in spinal fusion. METHODS:Silk fibroin/nano-hydroxyapatite composite was synthesized by the co precipitation method with silk fibroin and calcium phosphate as raw materials, to simulate the structure and composition of the bone tissue. The crystal phase composition and microstructure of the composite scaffold were analyzed by X-ray diffraction and transmission electron microscope. Osteoblasts were seeded onto the composite, and the cel adhesion and proliferation were observed under inverted microscope. The lumbar posterolateral spinal fusion models were established in the New Zealand rabbits, fol owed by treated with autologous bone graft (control group) and composite (composite group), respectively. The gross, radiological and histological observations of bone fusion were compared between groups. RESULTS AND CONCLUSION:Silk fibroin/nano-hydroxyapatite composite appeared to be fascicular under electron microscope, the length was 200-500 nm and width was 20-30 nm. And the hydroxyapatite was about 200 nm in length and 50 nm in width. X-ray diffraction showed that the bottom of diffraction peak was wide, and the peak was not sharp. Transmission electron microscope found that cel s adhered wel onto the composite scaffold at 2 days. Scanning electron microscope showed that the polygonal, oval or conical cel s covered most of the composite scaffold holes, presented obvious mitotic phase at 5 days. The third generation of MC3T3-E cel s tended to rise at first 3 days, and then decreased. The fusion site of L5-6 transverse process was strong, and non-bony fusion occurred. At the same time, numerous new bones were visible in the composite group. Hematoxylin-eosin staining showed a large number of cel aggregation, abundant osteoblasts surrounding cartilage, and the bone tissues were in a regular arrangement in the composite group. Moreover, irregular trabecular bone with medul ary cavity was found in the composite material. These results suggest that the silk fibroin/nano-hydroxyapatite composite with the similar structure and composition of natural bone can achieve satisfactory fusion effect in the rabbit lumbar posterolateral fusion.

Objective To investigate the effects of electromagnetic irradiation on the activity of protein kinase C(PKC)in the cerebellum of rabbit.Methods Thirty rabbits were randomly divided into control and electromagnetic irradi.ation group (including 0 hour,3 hours,12 hours,24 hours and 72 hours subgroups after irradiation),rabbits in the electromagnetic irradiation group were exposed to 65 mW/cm~2 S frequency range electromagnetic irradiation for 30 minutes,the rectal temperature was taken immediately after irradiation and the specific absorption rate(SAR)values were calculated,the activity of PKC the cerebellum was determined with improved TaKai method.Results The rectal temperature of rabbits increased 2.35 ℃ after irradiation,SAR value was 4.54 kcal/kg.PKC activity decreased significantly after irradiation.Conclusion After 30 min of electromagnetic irradiation at 65 mW/cm~2,obvious heating effect will be detected in the rabbit and PKC activity in the cerebellum decreased significantly.

Objective To observe the effect of sinomenine(SN) in vitro on nuclear factor-?B(NF-?B) DNA binding activity and nuclear translocation of synoviocytes in collagen-induced arthritis(CIA) rats and explore its antiinflammatory mechanisms.Methods The experimental model of CIA rats was used and synoviocytes were collected.Cells were divided into five groups:normal control,CIA,CIA+10 ?mol/L methotrexate(MTX),CIA+50 ?mol/L SN,CIA+500 ?mol/L SN.Nuclear translocation of NF-?B p65 subunit and NF-?B DNA binding activity of synoviocytes were investigated by fluorescence labelling laser confocal scanning microscopy and electrophoretic mobility shift assay(EMSA) respectively.Results Compared to normal control,significant nuclear translocation of NF-?B p65 subunit was observed and NF-?B DNA binding activity was increased in synoviocytes of CIA rats(P

BACKGROUND:Although the traditional surgical treatment can improve the symptoms of patients with senile osteoporotic vertebral fracture, the treatment easily produces bone graft fusion failure and pseudoarticulation formation and affects clinical effects. OBJECTIVE:To investigate biomechanical properties of anterior cervical pedicle screw and the effects on osteoporotic vertebral stability. METHODS:A total of 16 fresh cadaver cervical specimens contained 64 motion segments (C3-4, C4-5, C5-6 and C6-7). The 64 segments by the way of implantation were randomly divided into ordinary anterior locking screw fixation group and lower anterior vertebral pedicle screw system group (32 segments per group). The mechanical properties were determined on the biomechanical testing machine for each group. RESULTS AND CONCLUSION:(1) Biomechanics:Compared with the ordinary anterior locking screw fixation group, the maximum pul-out strength, screw path length, postoperative vertebral column height, the maximum surface strain, strain maximum and the range of maximum values were increased in the lower anterior vertebral pedicle screw system group (P<0.05). (2) Results suggest that compared with the ordinary anterior locking screw fixation group, lower cervical anterior pedicle screw required larger extraction force and was more stable for osteoporotic vertebrae.

OBJECTIVETo observe the effect of bone morphogenetic protein-7 (BMP-7) on aristolchic acid induced renal tubular epithelial cell trans-differentiation to look for new therapeutic approach for aristolchic acid nephropathy (AAN).

METHODSIn vitro cultured human proximal renal tubular epithelial cell line HK-2 cells were treated with different concentrations of BMP-7 (75 ng/mL, 150 ng/mL and 300 ng/mL) after trans-differentiation of the cells was induced by AA (10 microg/mL). Levels of alpha-SMA mRNA and protein expressions were detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blotting respectively.

RESULTSBMP-7 reversed the AA inducing alpha-SMA expressions in HK-2 cells in a dose-dependent manner.

CONCLUSIONBMP-7 can inhibit the trans-differentiation of human renal tubular epithelial cell induced by AA, thereby might be a new potential drug for AAN prevention and treatment.

Actins ; metabolism ; Aristolochic Acids ; adverse effects ; Bone Morphogenetic Protein 7 ; pharmacology ; Cell Line ; Cell Transdifferentiation ; drug effects ; Epithelial Cells ; cytology ; Humans ; Kidney Tubules, Proximal ; cytology

Ex-RAD (ON01210) is a novel and efficient anti-radiation drug with low toxicity developed by US army and Onconova Pharmaceuticals in recent years. The significant survival advantage and low toxicity of Ex-RAD have contributed to the approval by the FDA as an investigational new drug in December 2008. Meanwhile, the drug is currently in phase I clinical trials in humans. In this paper, the chemical structure, synthesis route, detection method and pharmacological action of Ex-RAD were reviewed, and the appli-cation prospect of Ex-RAD was also explored.

Objective To investigate the effect of Shensong Yangxin capsule on cardiac remodelling of myocardial infarction mouse model and the possible molecular mechanisms. Methods Adult male C57BL/6J mice were divided into sham operation group(n=10), model control group(n=20)and Shensong Yangxin group(n=20)according to random number table. Left anterior descending branch of coronary artery was ligated to establish myocardic infarction model in the model control group and Shensong Yangxin group. From the 2nd day after the surgery, Shensong Yangxin ( 400 mg . kg-1 ) was intragastrically administered, and the death rate of the mice was observed.Four weeks after the surgery, echocardiography was used to measure the cardiac function;myocardiac infarction area was detected by pathological staining;the expression levels of cardiac remodelling markers and extracellular matrix proteins were detected by RT-PCR. The possible molecular mechanisms were screened by Western blotting. Results As compared with the model control group, Shensong Yangxin significantly reduced the mortality after myocardial infarction in mice(P<0.05), as well as the myocardial infarct size(P<0.05).The mRNA expression levels of cardiac remodelling markers ANP, BNP, and β-MHC and the extracellular matrix proteins(collagenⅠ, collagen Ⅲ, CTGF, TGFβ) decreased significantly in the Shensong Yangxin group as compared with the model control group. Western blotting showed that Shensong Yangxin significantly decreased activation of smad3, and reduced expression level of smad4. Conclusion Shensong Yangxin attenuates cardiac remodelling after myocardial infarction and the mechanism may be related with blockage of smad signaling pathway.

Objective To study the characteristics of heterotopic heart transplantation by echocardiographyand evaluate the heart function and acute rejection.Methods Five male 49-61 years old patients with heterotopic heart transplantation were studied using echocardiography pre-,intra-,post～operation and at follow-up.The anastomoses of great vessels of recipient with donor heart,chamber size,left ventricular wall thickness with wallmotion.left ventricular ejection fraction(LVEF),valvular function,pulmonary artery pressure,pericardial and pleural effusion were observed.Results All patients were end-stage coronary artery disease with NYHA class of received heterotopic heart transplantation with coronary artery bypass grafting.Some patients received mitral valvuloplasty,ventricular aneurysmectomy and stem cell transplantation simultaneously.During the operation,intra-operation transesophageal echocardiography was used to monitor and evaluate the surgical procedure with chamber sizes,anastomoses,native with donor heart ventricular and valvular function,and air bubble in chambers.The cardiac functions of donor heart backed to normal post operation,while the functions of recipient heart were increased gradually.All patients survived in 6 months and one year follow-up,with significantly improved functions of heart in situ[LVEF(44.0±15.6)%].The main reject reaction was small pericardial perfusion and pleural effusion.Conclusions Heterotopic heart transplantation has distinctive characteristics with echocardiography study,which is also an important technique in evaluation at pre-,intra-,post-operation and also at follow-up.

Purpose To investigate human soluble TRAIL(sTRAIL)protein expression and purification and its potential anti-tumor activity on hepatocellular carcinoma(HepG2).Methods Soluble TRAIL gene ligated with expression vector pPIC9 was transfected into GS115(his4)and the recombination strain expressing sTRAIL was screened by MD plate.The effects of different media,methanol inducement period,methanol concentration,and pH were investigated and optimized using shaking flask.The anti-tumor activity of sTRAIL with HepG2 cells Was analyzed after purification.Results The highest expression of sTRAIL was obtained at pH 6.0,1% methanol in BMMY medium,with the concentration of(58.7±2.4)mg/L at 48 h.Recombinant sTRAIL protein could induce HepG2 cells apoptosis and inhibit HepG2 cells proliferation effectively.Conclusion The optimized condition of human sTRAIL expression and purification Was developed and the obtained recombinant sTRAIL protein may be a promising therapeutic agent for hepatocellular carcinoma.