Human sTNFR1 (soluble tumor necrosis factor receptor 1) gene was amplified by RT-PCR from Hela cells. A recombinant expression vector of sTNFR1-MBP was constructed in pMAL-c2x, and transformed into E. Coli JM109.It was sequenced and confirmed to be identifical to the sTNFR1 gene in data bank. Recombinant protein sTNFR1-MBP was induced by IPTG and purified by Amylose resin Affinity Chromatography. sTNFR1-MBP was binded to sTNFR1's antibody in Western-blotting. From MTT assays, the results showed that sTNFR1-MBP could effectively block the cytotoxicity mediated by TNF?on QSG7701 cells. Annexin V-FITC staining and flowcytometry were used to observe the recombinant protein's anti-apoptosis capacity and the recombinant protein has marked anti-apoptosis effect in vitro.sTNFR1-MBP had good biological activity and it will be employed in further study.

OBJECTIVETo review the recent developments in the mechanisms of glucocorticoids induced osteonecrosis of femoral head (ONFH) and introduce a new theory of ONFH.

DATA SOURCESBoth Chinese- and English-language literatures were searched using MEDLINE (1997 - 2011), Pubmed (1997 - 2011) and the Index of Chinese-language Literature (1997 - 2011).

STUDY SELECTIONData from published articles about mechanisms of glucocorticoids induced ONFH in recent domestic and foreign literature were selected. Data extraction Data were mainly extracted from 61 articles which are listed in the reference section of this review.

RESULTSGlucocorticoids are steroid hormones secreted by the adrenal cortex that play a pivotal role in the regulation of a variety of developmental, metabolic and immune functions. However, high dose of exogenous glucocorticoids usage is the most common non-traumatic cause of ONFH. Glucocorticoids can affect the metabolisms of osteoblasts, osteoclasts, bone marrow stromal cells and adipocytes which decrease osteoblasts formation but increase adipocytes formation and cause ONFH finally.

CONCLUSIONSGlucocorticoids affect the differentiation of mesenchymal stem cells, through activating or inhibiting the related transcript regulators of osteogenesis and adipogenesis. At last, the size and volume of mesenchymal stem cells derived adipocytes will increase amazingly, but the osteoblasts will be decreased obviously. In the meantime, the activity of the osteoclasts will be activated. So, these mechanisms work together and lead to ONFH.

Animals ; Bone Marrow Cells ; drug effects ; metabolism ; Femur Head ; cytology ; Glucocorticoids ; metabolism ; pharmacology ; Humans ; Osteonecrosis ; etiology ; metabolism ; Stromal Cells ; drug effects ; metabolism

OBJECTIVETo observe the therapeutic effect of the low molecular weight heparin calcium (LMWHC) combined with Yinxin Damo Injection (YDI) on the prevention of deep venous thrombosis (DVT) after the operation of intertrochanteric fracture.

METHODSSixty-two patients with intertrochanteric fracture were randomly assigned to the control group and the combined group, 31 in each group. Patients in the control group were subcutaneously injected with 6 000 IU LMWHC, twice daily. On this basis of the aforesaid treatment, patients in the combined group were intravenously dripped with YDI at the daily dose of 20 mL, once daily. The treatment lasted for 10 days in both groups. The DVT incidence rate, postoperative drainage amount of blood, activated partial thromboplastin time (APTT), plasma prothrombin time (PT), platelet count, and the maximal swelling rate of lower limbs after surgery were compared between the two groups after operation.

RESULTSIn the control group DVT occurred in 5 cases (16. 1%) while it occurred in 2 cases (6.5%) in the combined group, showing significant differences (P < 0.05). Both APTT and PT were shortened in the two groups on the 2nd day after medication (P < 0.01). Compared with the control group, it was more obvious in the combined group, showing statistical significance (P < 0.05). Both APTT and PT were shortened in the control group on the 10th day after medication when compared with before treatment (P < 0.01). But they were postponed in the combined group when compared with before medication, showing statistical difference when compared with the control group (P < 0.05). The platelet count increased in the combined group on the 10th day after medication. Besides, it was higher than that in the control group, showing statistical difference (P < 0.05). The platelet counts were in the normal range.

CONCLUSIONSLMWSC combined with YDI could not only reduce the incidence of DVT after the operation of intertrochanteric fracture, but also change the state of higher blood coagulation. It showed significant effect on preventing DVT after the operation of intertrochanteric fractures.

Aged ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Ginkgo biloba ; Heparin, Low-Molecular-Weight ; therapeutic use ; Hip Fractures ; surgery ; Humans ; Injections ; Lower Extremity ; blood supply ; Male ; Middle Aged ; Postoperative Complications ; prevention & control ; Venous Thrombosis ; prevention & control

Objective To observe the effect of T-2 toxin on growth and development of rat epiphyseal plate of left knee and metaphyseal bone of femur and tibia in normal and low nutritional status, to find out possible pathogenic factors of Kashin-Beck disease and provide experimental basis for early intervention. Methods Ninety 3-week-old Wistar rats, weighing 60 - 70 g, were randomly divided into three groups: control group(general feed), T-2 toxin + general feed group, T-2 toxin + low nutrition feed group, thirty rats in each group with equally sex ratio. T-2 toxin (1.0 mg/kg) was administered orally 5 times a week via a gavage needle for 4 weeks. The change of hair, activity and body weight was observed. After 1, 2, 4 weeks, the epiphyseal plate of left knee and metaphyseal bone of femur and tibia (including distal femur and proximal tibia) were collected. Specimens were processed with HE and Masson staining. The morphology of chondrocytes and matrix collagen content in epiphyseal plate was observed. Trabecular bone volume fraction in tibial metaphyseal bone was analyzed by Image-Pro Plus 6.0 software. Results In the control group, rats were in good movement and hair with light, but in T-2 toxin + general feed group and T-2 toxin + low nutrition feed group, rats were found with reduced activities and hair with dark color. Body weights(g) of the control group, the T-2 toxin + general feed group and the T-2 toxin + low nutrition feed group were 81.0 ± 6.2, 79.0 ±5.1, 77.0 ± 7.5, respectively, by the end of first week; 101.8 ± 6.7, 97.0 ± 6.8, 93.0 ± 5.3, respectively, by the end of second week; 151.1 ± 15.7, 126.5 ± 11.9, 106.5 ± 11.5, respectively, by the end of fourth week. There was significant difference in groups by second week and the fourth week (F = 9.72, 41.65, all P ＜ 0.05 ). There was significant difference among multi-groups by the fourth week(all P ＜ 0.01 ). Under light microscope, at the second weeks, coagulative necrosis of chondrocytes was found in hypertrophic zone in the two groups with T-2 toxin; at the fourth weeks, cell necrosis increased. Masson staining showed collagen staining in the two groups with T-2 toxin significantly turned to clear pale coloration, indicating that the collagen matrix was significantly reduced. Image analysis showed there was significant difference in groups at the second and fourth week(F= 9.72, 41.65, all P＜ 0.05)in tibial metaphyseal trabecular bone volume fraction. There was significant difference between T-2 toxin + low nutrition feed group[(0.55 ± 0.12)％, (0.21 ± 0.0)％] and control group[(0.67 ± 0.09)％, (0.51 ± 0.14)％] by the second and fourth week(all P ＜ 0.01 ). Conclusions Under normal nutritional status, T-2 toxin can induce hypertrophic epiphyseal cartilage necrosis, collagen content decreased in epiphyseal plate, metaphyseal trabecular bone formation disorders; in the low nutritional status, T-2 toxin can lead to rat epiphyseal necrosis and significant metaphyseal bone disorder, but whether the performance is related to Kaschin-Beck disease needs to be studied further.

AIMTo prepare the sustained-release nitrendipine microspheres with a solid dispersed structure in liquid system.

METHODSThe sustained-release nitrendipine microspheres with a solid dispersed structure was prepared in liquid system by combining spherical crystallization technique and solvent deposition method in one step. The resultant microspheres were evaluated for the recovery, micromeritc properties, incorporation efficiency. The factors of effect on the formation and the release rate of microspheres were also investigated.

RESULTSThe recovery of microspheres (280-900 microns) was more than 70% and the bulk density was around 0.7 kg.L-1. The incorporation efficiency always exceeded 95%. The formation of microspheres was mainly affected by the amount of bridging liquid and the emulsifying agents in poor solvent. The release rate of nitrendipine from the microspheres could be controlled as desired by adjusting the ratio of talc to Eudragit RS PO in the formulation.

CONCLUSIONThe presented method was suitable for preparing sustained-release microspheres of a water insoluble drug.

Delayed-Action Preparations ; Drug Carriers ; Microspheres ; Nitrendipine ; administration & dosage ; Particle Size ; Technology, Pharmaceutical ; methods

OBJECTIVETo investigate the clinical results of modified Sutherland pelvic osteotomy for developmental dysplasia of hip (DDH).

METHODSSutherland pelvic osteotomy were performed in 10 patients (11 hips) with DDH. Among them, there were 3 male (3 hips) and 7 female (8 hips) patients, aged (32 ± 8) years. During operation, arthroscopes were performed additionally to remove the existing hyperplasia tissue in the fossae ovalis and trimming acetabulum and glenoid labrum, thus to insure the better match between the femoral head and the realigned acetabulum. The change of imaging indexes were acquired by comparing the preoperative X-ray with the postoperative X-ray. The change of hip function and life quality were acknowledged according to contrast and analysis Harris hip score and Short Form 12-items Health Survey (SF-12) before and after osteotomy.

RESULTSAll patients were followed up for a mean of (5.2 ± 2.3) years, the osteotomy were all union in 3 months. The acetabular head index was 71 ± 8 before operation, and 86 ± 4 after operation. The pre- and post-operative centre edge angle were (7 ± 9)° and (33 ± 9)°, sharp angle were (48 ± 4)° and (37 ± 5)°, acetabular index angle were (24 ± 8)° and (11 ± 5)° respectively. The average Harris score improved from 42 ± 13 preoperatively to 90 ± 5 postoperatively, with 100% excellent and good results. Every domains of SF-12 was improved in the different extents postoperatively, the improvement of physical component summary was more conspicuous than mental component summary. The imaging indexes, Harris and SF-12 were all improved with significant difference (all P < 0.05).

CONCLUSIONSThe modified Sutherland pelvic osteotomy is effective. It could increase the load bearing capacity of hip, and improve the quality of life.

Adult ; Arthroscopy ; Female ; Hip Dislocation, Congenital ; surgery ; Humans ; Male ; Osteotomy ; methods ; Pelvic Bones ; surgery ; Treatment Outcome

Apoptosis ; drug effects ; HeLa Cells ; Hepatocytes ; cytology ; drug effects ; Humans ; Receptors, Tumor Necrosis Factor, Type I ; genetics ; Transfection ; Tumor Necrosis Factor-alpha ; pharmacology

OBJECTIVEA proteomics approach was applied for finding multi-drug resistance (MDR) related proteins in hepatic cancer cell lines.

METHODSTwo-dimensional electrophoresis (2-DE) was used to separate the total proteins of vincristine-resistant hepatic cancer cell line HepG2/VCR and its counterpart HepG2. The differential expression proteins between the two cell lines were identified by both MALDI-TOF-MS and ESI-Q-TOF-MS. Heat-shock protein 27 (HSP27), one of the differential expression proteins in the development of MDR of HepG2/VCR, was analyzed by HSP27 antisense oligonucleotides (ASOs).

RESULTSAs a high expression protein in HepG2/VCR, HSP27 was identified, and the suppression of HSP27 expression by HSP27 ASO enhanced the vincristine chemosensitivity of HepG2/VCR (P less than 0.05).

CONCLUSIONHSP27 is linked to MDR in human hepatic cancer cell line HepG2/VCR.

Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; HSP27 Heat-Shock Proteins ; metabolism ; Hep G2 Cells ; Humans ; Liver Neoplasms ; metabolism

Adult ; Aged ; External Fixators ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Male ; Middle Aged ; Tibial Fractures ; surgery

OBJECTIVETo evaluate clinical and radiographic results and influencing factors of hybrid total hip arthroplasty (THA).

METHODSTotally 126 patients (135 hips) from January 1999 to December 2001 accepted hybrid THA were followed up. Components migration, periprosthetic bone changes, the polyethylene wear rate were measured radiologically. Kaplan-Meier analysis was performed to evaluate the survival of the acetabular and femoral components. End point was obvious radiological loosening or revision either or both of the acetabular and femoral components for aseptic loosening, infection or osteolysis debridement surgery.

RESULTSA total of 79 cases (85 hips) had been given follow-up. Using revision as the end point, the survival rate of acetabular was 95.2%, cemented femoral components was 98.8%. Using loosening as the end point for failure, the survival rate of acetabular was 97.6%, cemented femoral components was 100%.

CONCLUSIONSThe hybrid total hip prosthesis long-term survival rate was satisfactory, especially in the femoral side. As a result of third-cement technology, the long-term survival rate of the femoral components was close to the modern cementless prosthesis.

Adult ; Aged ; Aged, 80 and over ; Arthroplasty, Replacement, Hip ; methods ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome